pCDH- CMV- MCS- EF1- CopGFP- T2A- Puro

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  • Model: PVT2333
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pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro

PVT2333     2ug

 

pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro Information

Promoter: EF-1 alpha core, CMV
Replicator: pUC ori, SV40 ori
Terminator: SV40 poly (A) signal
Plasmid classification: virus series, lentivirus cloning vector
Plasmid size: 8189bp
Prokaryotic resistance: Amp
Screening markers: Puro
Cloned strain: Stbl3
Culture conditions: 37 centigrade, aerobic LB
Expression host: mammalian cells
Induction mode: no induction, instantaneous expression
5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'sequencing primers: primers designed according to sequence
Use:Lentiviral

 

pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro Description

PCDH-CMV-MCS-EF1-CopGFP-T2A-Puro polyclonal site (MCS) - in MCS is located in the CMV promoter downstream of interest gene cloning.To improve stability and CMV driven transcription translation. SV40 multi - poly adenosine acidification signal to enable the efficiency of transcription and transcription termination. The hybrid / RSV 5ltr promoter provides a high level of viral transcript expression in 293 cells of this full-length producer.

pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro-polyclonal-site

 

pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro  Sequence

LOCUS       Exported                8189 bp ds-DNA     circular SYN 30-AUG-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 2

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8189)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4

FEATURES             Location/Qualifiers

     source          1..8189

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     LTR             234..414

                     /note="5' LTR (truncated)"

                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"

     misc_feature    458..583

                     /note="HIV-1 Psi"

                     /note="packaging signal of human immunodeficiency virus 

                     type 1"

     misc_feature    1076..1309

                     /note="RRE"

                     /note="The Rev response element (RRE) of HIV-1 allows for 

                     Rev-dependent mRNA export from the nucleus to the 

                     cytoplasm."

     misc_feature    1798..1915

                     /note="cPPT/CTS"

                     /note="central polypurine tract and central termination 

                     sequence of HIV-1"

     promoter        2005..2208

                     /note="CMV promoter"

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     promoter        2348..2559

                     /note="EF-1-alpha core promoter"

                     /note="core promoter for human elongation factor 

                     EF-1-alpha"

     LTR             2572..2840

                     /note="5' LTR (truncated)"

                     /note="truncated 5' long terminal repeat (LTR) from human 

                     T-cell leukemia virus (HTLV) type 1"

     CDS             2899..3561

                     /codon_start=1

                     /product="green fluorescent protein 2 from Pontellina 

                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"

                     /note="CopGFP"

                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT

                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY

                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG

                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"

     CDS             3631..3684

                     /codon_start=1

                     /product="2A peptide from Thosea asigna virus capsid 

                     protein"

                     /note="T2A"

                     /note="Eukaryotic ribosomes fail to insert a peptide bond 

                     between the Gly and Pro residues, yielding separate 

                     polypeptides."

                     /translation="EGRGSLLTCGDVEENPGP"

     CDS             3685..4284

                     /codon_start=1

                     /gene="pac from Streptomyces alboniger"

                     /product="puromycin N-acetyltransferase"

                     /note="PuroR"

                     /note="confers resistance to puromycin"

                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER

                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA

                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS

                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"

     misc_feature    4285..4872

                     /note="WPRE"

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     CDS             complement(4756..4767)

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /translation="IEGR"

     LTR             4946..5179

                     /note="3' LTR (Delta-U3)"

                     /note="self-inactivating 3' long terminal repeat (LTR) from

                     HIV-1"

     polyA_signal    5251..5372

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

     rep_origin      5391..5526

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     primer_bind     complement(5564..5580)

                     /note="M13 rev"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

     protein_bind    5588..5604

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="lac operator"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     promoter        complement(5612..5642)

                     /note="lac promoter"

                     /note="promoter for the E. coli lac operon"

     protein_bind    5657..5678

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding site"

                     /note="CAP binding activates transcription in the presence 

                     of cAMP."

     rep_origin      complement(5966..6554)

                     /direction=LEFT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(6725..7585)

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /note="AmpR"

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     promoter        complement(7586..7690)

                     /gene="bla"

                     /note="AmpR promoter"

     primer_bind     8164..8180

                     /note="M13 fwd"

                     /note="common sequencing primer, one of multiple similar 

                     variants"

ORIGIN

        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca

       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta

      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga

      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc

      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta

      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact

      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg

      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct

      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt

      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag

      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt

      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt

      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg

      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag

      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag

      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg

      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag

     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag

     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc

     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga

     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc

     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg

     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata

     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa

     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga

     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa

     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat

     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt

     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg

     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt

     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc

     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga

     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac

     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga

     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg

     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg

     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac

     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc

     2281 tagcgaattc gaatttaaat cggatccgcg gccgcaagga tctgcgatcg ctccggtgcc

     2341 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc

     2401 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac

     2461 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg

     2521 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc

     2581 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc

     2641 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta

     2701 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc

     2761 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct

     2821 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag

     2881 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac

     2941 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc

     3001 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg

     3061 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg

     3121 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc

     3181 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag

     3241 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc

     3301 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc

     3361 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac

     3421 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc

     3481 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc

     3541 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc

     3601 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt

     3661 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc

     3721 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc

     3781 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc

     3841 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg

     3901 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc

     3961 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc

     4021 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc

     4081 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag

     4141 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag

     4201 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc

     4261 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg

     4321 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct

     4381 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg

     4441 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact

     4501 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc

     4561 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc

     4621 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag

     4681 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc

     4741 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg

     4801 gctctgcggc ctcttccgcg tctccgcctt cgccctcaga cgagtcggat ctccctttgg

     4861 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc

     4921 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat

     4981 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct

     5041 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta

     5101 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca

     5161 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt

     5221 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac

     5281 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt

     5341 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc

     5401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg

     5461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg

     5521 gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg

     5581 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta

     5641 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg

     5701 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga

     5761 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg

     5821 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag

     5881 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc

     5941 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca

     6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt

     6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc

     6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc

     6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc

     6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact

     6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg

     6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta

     6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca

     6481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa

     6541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg

     6601 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc

     6661 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg

     6721 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat

     6781 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg

     6841 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa

     6901 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca

     6961 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc

     7021 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt

     7081 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa

     7141 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat

     7201 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct

     7261 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga

     7321 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag

     7381 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga

     7441 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca

     7501 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg

     7561 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc

     7621 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag

     7681 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca

     7741 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg

     7801 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag

     7861 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg

     7921 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg

     7981 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc

     8041 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga

     8101 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac

     8161 gttgtaaaac gacggccagt gccaagctg

//

 

Caution:
Product is for research use only!

 

 

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pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro,Plasmid pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro,pCDH-CMV-MCS-EF1-CopGFP-T2A-Puro vector

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