pBIND

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  • Model: PVT1701
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pBIND

PVT1701        2ug
 

pBIND Informaiton

Promoter: T7, T3, CMV

Replicator: pUC ori, F1 ori, SV40 ori

Terminator: SV40 poly (A) signal

Plasmid classification: mammalian, mammalian two hybrid vectors

Plasmid size: 6360bp

Prokaryotic resistance: Amp

Cloned strain: DH5 alpha

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

 

pBIND Description

PBIND is the yeast GAL4 gene of the high copy plasmid CMV immediate early promoter in which the gene expression of the driving gene is expressed in part (1 - 147 of amino acids) containing the DNA binding domain. The influencing DNA binding domain sequence is carried by a chimeric flanked polyclonal region, an intron, a termination codon, and a polyadenylation SV40 late region one. The T7 and T3 flanked regions of the fusion gene are products of the target synthesizing sense and antisense RNA, respectively, by the initiator of the RNA polymerase. The carrier of luciferase gene is the intron and growth hormone of the early promoter of this preceded SV40 gene. The increased protein expression of mRNA introns can be stabilized through transport to the cytoplasm and nucleus (9 – 12). The polyadenylate sequence was synthesized by 3 of the sleeping kidney luciferase gene. The backbone contains a selection of single-stranded DNA (the list of beta-lactamase genes in Escherichia coli) from the origin and production of F1 plasmid replication.

pBIND-vector

 

pBIND Sequence

LOCUS       Exported                6360 bp ds-DNA     circular SYN 05-SEP-2016

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    Untitled 8

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 6360)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Monday, September 5, 2016 from SnapGene Viewer 3.1.4

FEATURES             Location/Qualifiers

     source          1..6360

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        138..517

                     /note="CMV enhancer"

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        518..729

                     /note="CMV promoter"

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     intron          890..1022

                     /note="chimeric intron"

                     /note="chimera between introns from human beta-globin and 

                     immunoglobulin heavy chain genes"

     promoter        1067..1085

                     /note="T7 promoter"

                     /note="promoter for bacteriophage T7 RNA polymerase"

     CDS             1116..1556

                     /codon_start=1

                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"

                     /product="DNA binding domain of the GAL4 transcriptional 

                     activator"

                     /note="GAL4 DNA binding domain"

                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK

                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK

                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"

     promoter        complement(1640..1657)

                     /note="T3 promoter"

                     /note="promoter for bacteriophage T3 RNA polymerase"

     polyA_signal    1675..1796

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

     rep_origin      1982..2437

                     /direction=RIGHT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        2509..2866

                     /note="SV40 promoter"

                     /note="SV40 enhancer and early promoter"

     rep_origin      2717..2852

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     CDS             3208..4143

                     /codon_start=1

                     /product="luciferase from the anthozoan coelenterate 

                     Renilla reniformis (sea pansy)"

                     /note="Rluc"

                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN

                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW

                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI

                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE

                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV

                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"

     polyA_signal    4201..4249

                     /note="synthetic polyadenylation signal"

     promoter        4563..4667

                     /gene="bla"

                     /note="AmpR promoter"

     CDS             4668..5528

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /note="AmpR"

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

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      5699..6287

                     /direction=RIGHT

                     /note="ori"

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

                     replication"

ORIGIN

        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta

       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc

      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg

      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc

      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat

      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc

      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga

      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg

      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac

      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt

      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg

      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata

      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac

      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt

      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa

      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact

      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac

     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact

     1081 ataggctagc cagcttgaag caagcctcct gaaagatgaa gctactgtct tctatcgaac

     1141 aagcatgcga tatttgccga cttaaaaagc tcaagtgctc caaagaaaaa ccgaagtgcg

     1201 ccaagtgtct gaagaacaac tgggagtgtc gctactctcc caaaaccaaa aggtctccgc

     1261 tgactagggc acatctgaca gaagtggaat caaggctaga aagactggaa cagctatttc

     1321 tactgatttt tcctcgagaa gaccttgaca tgattttgaa aatggattct ttacaggata

     1381 taaaagcatt gttaacagga ttatttgtac aagataatgt gaataaagat gccgtcacag

     1441 atagattggc ttcagtggag actgatatgc ctctaacatt gagacagcat agaataagtg

     1501 cgacatcatc atcggaagag agtagtaaca aaggtcaaag acagttgact gtatcgccgg

     1561 aattcccggg gatccgtcga cttgacgcgt tgatatcatc tagagcggcc gcaggtacct

     1621 gaataactaa ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat

     1681 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg

     1741 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca

     1801 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa

     1861 gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa

     1921 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg

     1981 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc

     2041 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttcacgcc

     2101 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt

     2161 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg

     2221 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt

     2281 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta

     2341 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt

     2401 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta

     2461 cgcatctgtg cggtatttca caccgcatac gcggatcttc cgtaccttct gaggcggaaa

     2521 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg

     2581 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg

     2641 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc

     2701 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca

     2761 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt

     2821 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttgattcttc

     2881 tgacacaaca gtctcgaact taaggctaga attctggcta caggtaagcg cccctaaaat

     2941 ccctttgggc acaatgtgtc ctgaggggag aggcagcgac ctgtagatgg gacgggggca

     3001 ctaaccctca ggtttggggc ttctgaatgt gagtatcgcc atgtaagccc agtatttggc

     3061 caatctcaga aagctcctgg tccctggagg gatggagaga gaaaaacaaa cagctcctgg

     3121 agcagggaga gtgctggcct cttgctctcc ggctccctct gttgccctct ggtttctccc

     3181 caggctcccg gacgtcctct agccaccatg acttcgaaag tttatgatcc agaacaaagg

     3241 aaacggatga taactggtcc gcagtggtgg gccagatgta aacaaatgaa tgttcttgat

     3301 tcatttatta attattatga ttcagaaaaa catgcagaaa atgctgttat ttttttacat

     3361 ggtaacgcgg cctcttctta tttatggcga catgttgtgc cacatattga gccagtagcg

     3421 cggtgtatta taccagacct tattggtatg ggcaaatcag gcaaatctgg taatggttct

     3481 tataggttac ttgatcatta caaatatctt actgcatggt ttgaacttct taatttacca

     3541 aagaagatca tttttgtcgg ccatgattgg ggtgcttgtt tggcatttca ttatagctat

     3601 gagcatcaag ataagatcaa agcaatagtt cacgctgaaa gtgtagtaga tgtgattgaa

     3661 tcatgggatg aatggcctga tattgaagaa gatattgcgt tgatcaaatc tgaagaagga

     3721 gaaaaaatgg ttttggagaa taacttcttc gtggaaacca tgttgccatc aaaaatcatg

     3781 agaaagttag aaccagaaga atttgcagca tatcttgaac cattcaaaga gaaaggtgaa

     3841 gttcgtcgtc caacattatc atggcctcgt gaaatcccgt tagtaaaagg tggtaaacct

     3901 gacgttgtac aaattgttag gaattataat gcttatctac gtgcaagtga tgatttacca

     3961 aaaatgttta ttgaatcgga cccaggattc ttttccaatg ctattgttga aggtgccaag

     4021 aagtttccta atactgaatt tgtcaaagta aaaggtcttc atttttcgca agaagatgca

     4081 cctgatgaaa tgggaaaata tatcaaatcg ttcgttgagc gagttctcaa aaatgaacaa

     4141 taattctagc cctgaataag tgataataag cggatgaatg gcagaaattc gtcgaagcgc

     4201 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg

     4261 ataaggatcg gaagatccgc gtatggtgca ctctcagtac aatctgctct gatgccgcat

     4321 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc

     4381 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt

     4441 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat

     4501 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg

     4561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga

     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac

     4681 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc

     4741 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca

     4801 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc

     4861 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg

     4921 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac

     4981 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca

     5041 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg

     5101 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac

     5161 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg

     5221 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat

     5281 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg

     5341 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg

     5401 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc

     5461 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc

     5521 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt

     5581 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt

     5641 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt

     5701 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag

     5761 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca

     5821 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca

     5881 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg

     5941 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg

     6001 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct

     6061 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga

     6121 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc

     6181 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg

     6241 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg

     6301 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct

//

Caution:
Product is for research use only!

 

 

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