pCIG

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  • Model: PVT10732
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pCIG

PVT10732   2ug

 

pCIG Description
PCIG plasmid is an over expression vector of mammalian cells. CAG promoter drives the expression of target genes.

 

pCIG Information

Promoter: CAG promoter

Replicon: Ori, SV40 ori

Terminator: beta -globin poly (A) signal

Plasmid classification: lactation cell plasmid; lactation expression plasmid; pCAG series plasmid.

Plasmid size: 6200bp

Prokaryotic resistance: ampicillin Amp

Selection marker: EGFP

Cloning strains: E. coli DH5 and E.

Culture conditions: 37 C, aerobic, LB

Expression host: mammalian cells such as 293T

 

pCIG Sequence

LOCUS       Exported                6200 bp ds-DNA    circular SYN 18-AUG-2017

EYWORDS    pCIG

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6200)

FEATURES             Location/Qualifiers

     source          1..6200

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      complement(42..630)

                     /direction=LEFT

                     /note="ori"

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

                     replication"

     CDS             complement(801..1661)

                     /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(1662..1766)

                     /gene="bla"

                     /note="AmpR promoter"

     enhancer        1797..2176

                     /note="CMV enhancer"

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        2178..2455

                     /note="chicken beta-actin promoter"

     intron          2456..3468

                     /note="chimeric intron"

                     /note="chimera between introns from chicken beta-actin and 

                     rabbit beta-globin"

     misc_feature    3594..4172

                     /note="IRES2"

                     /note="internal ribosome entry site (IRES) of the 

                     encephalomyocarditis virus (EMCV)"

     CDS             4173..4892

                     /codon_start=1

                     /product="enhanced GFP"

                     /note="EGFP"

                     /note="mammalian codon-optimized"

                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL

                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD

                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK

                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL

                     EFVTAAGITLGMDELYK"

     polyA_site      5049..5104

                     /note="beta-globin poly(A) signal"

                     /note="rabbit beta-globin polyadenylation signal"

     promoter        5668..5863

                     /note="SV40 promoter"

                     /note="SV40 early promoter"

     rep_origin      5714..5849

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     polyA_signal    5869..6003

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

ORIGIN

        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc

       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta

      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg

      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc

      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac

      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac

      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg

      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga

      481 agnacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt

      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag

      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct

      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg

      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat

      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc

      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg

      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct

      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca

     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg

     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg

     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc

     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag

     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg

     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag

     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat

     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg

     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca

     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca

     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat

     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag

     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca

     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata

     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga

     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt

     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt

     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca

     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt

     2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc

     2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg

     2281 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg

     2341 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt

     2401 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt

     2461 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct

     2521 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg

     2581 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa

     2641 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg

     2701 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg

     2761 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc

     2821 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg

     2881 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc

     2941 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg

     3001 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct

     3061 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga

     3121 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt

     3181 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc

     3241 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt

     3301 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac

     3361 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc

     3421 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac

     3481 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgaggt cgatcgacgg

     3541 tatcgataag cttgatatcg aattcctgca gcccggccgc tacgtaaatt ccgccccccc

     3601 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat

     3661 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt

     3721 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt

     3781 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc

     3841 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc

     3901 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat

     3961 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc

     4021 ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg

     4081 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct

     4141 ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca

     4201 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg

     4261 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca

     4321 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc

     4381 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc

     4441 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc

     4501 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg

     4561 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca

     4621 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc

     4681 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg

     4741 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca

     4801 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga

     4861 tcactctcgg catggacgag ctgtacaagt aaagcggccg caattcactc ctcaggtgca

     4921 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg

     4981 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac

     5041 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct

     5101 atcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt

     5161 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag

     5221 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt

     5281 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct

     5341 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt

     5401 catagctgtc cctcttctct tatggagatc cctcgacctg caccgtcgac cagctggtcg

     5461 acggtgcacc gtcgaccagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt

     5521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg

     5581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt

     5641 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc

     5701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg

     5761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca

     5821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt

     5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt

     5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg

     6001 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct

     6061 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat

     6121 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga

     6181 acatgtgagc aaaaggccag

//

 

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