LOCUS       V019063                 9603 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019063
VERSION     V019063
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        50..1228
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             1281..4274
                     /label="GTF2I(NM_032999)"
                     /note="GTF2I(NM_032999)"
                     /gene="GTF2I"
     CDS             4290..4355
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4379..4967
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5033..5081
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5283..5711
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5725..6054
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6121..6915
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    7091..7212
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7261..7277)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7285..7301
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7309..7339)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7354..7375
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7663..8251)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8422..9282)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(9283..9387)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg gctccggtgc
       61 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      121 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      181 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      241 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      301 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
      361 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      421 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      481 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      541 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      601 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      661 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      721 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      781 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
      841 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
      901 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
      961 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1021 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1081 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1141 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1201 tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactggcta gcgtttaaac
     1261 ttaagcttgg taccgccacc atggcccaag ttgcaatgtc caccctcccc gttgaagatg
     1321 aggagtcctc ggagagcagg atggtggtga cattcctcat gtcagctctc gagtccatgt
     1381 gtaaagaact ggccaagtcc aaagccgaag tggcctgcat tgcagtgtat gaaacagacg
     1441 tgtttgtcgt cggaactgaa agaggacgtg cttttgtcaa taccagaaag gattttcaaa
     1501 aagattttgt aaaatattgt gttgaagaag aagaaaaagc tgcagagatg cataaaatga
     1561 aatctacaac ccaggcaaat cggatgagtg tagatgctgt agaaattgaa acactcagaa
     1621 aaacagttga ggactatttc tgcttttgct atgggaaagc tttaggcaaa tccacagtgg
     1681 tacctgtacc atatgagaag atgctgcgag accagtcggc tgtggtagtg caggggcttc
     1741 cggaaggtgt tgcctttaaa caccccgaga actatgatct tgcaaccctg aaatggattt
     1801 tggagaacaa agcagggatt tcattcatca ttaagagacc ttttttagag ccaaagaagc
     1861 atgtaggtgg tcgtgtgatg gtaacagatg ctgacaggtc aatactatct ccaggtggaa
     1921 gttgtggccc catcaaagtg aaaactgaac ccacagaaga ttctggtatt tccctggaaa
     1981 tggcagctgt gacagtaaag gaagaatcag aagatcctga ttattatcaa tataacattc
     2041 aagcaggccc ttctgaaact gatgatgttg atgaaaaaca gcccctatcg aagcctttgc
     2101 aaggaagcca ccattcttca gagggcaatg aaggcacaga aatggaagta ccagcagaag
     2161 attctactca acatgtccct tcagaaacaa gtgaggaccc tgaagttgag gtgactattg
     2221 aagatgatga ttattctcca ccgtctaaga gaccaaaggc caatgagcta ccgcagccac
     2281 cagtcccgga acccgccaat gctgggaagc ggaaagtgag ggagttcaac ttcgagaaat
     2341 ggaatgctcg catcactgat ctacgtaaac aagttgaaga attgtttgaa aggaaatatg
     2401 ctcaagccat aaaagccaaa ggtccggtga cgatcccgta ccctcttttc cagtctcatg
     2461 ttgaagatct ttatgtagaa ggacttcctg aaggaattcc ttttagaagg ccatctactt
     2521 acggaattcc tcgcctggag aggatattac ttgcaaagga aaggattcgt tttgtgatta
     2581 agaaacatga gcttctgaat tcaacacgtg aagatttaca gcttgataag ccagcttcag
     2641 gagtaaagga agaatggtat gccagaatca ctaaattaag aaagatggtg gatcagcttt
     2701 tctgcaaaaa atttgcggaa gccttgggga gcactgaagc caaggctgta ccgtaccaaa
     2761 aatttgaggc acacccgaat gatctgtacg tggaaggact gccagaaaac attcctttcc
     2821 gaagtccctc atggtatgga atcccaaggc tggaaaaaat cattcaagtg ggcaatcgaa
     2881 ttaaatttgt tattaaaaga ccagaacttc tgactcacag taccactgaa gttactcagc
     2941 caagaacgaa tacaccagtc aaagaagatt ggaatgtcag aattaccaag ctacggaagc
     3001 aagtggaaga gatttttaat ttgaaatttg ctcaagctct tggactcacc gaggcagtaa
     3061 aagtaccata tcctgtgttt gaatcaaacc cggagttctt gtatgtggaa ggcttgccag
     3121 aggggattcc cttccgaagc cctacctggt ttggaattcc acgacttgaa aggatcgtcc
     3181 gcgggagtaa taaaatcaag ttcgttgtta aaaaacctga actagttatt tcctacttgc
     3241 ctcctgggat ggctagtaaa ataaacacta aagctttgca gtcccccaaa agaccacgaa
     3301 gtcctgggag taattcaaag gttcctgaaa ttgaggtcac cgtggaaggc cctaataaca
     3361 acaatcctca aacctcagct gttcgaaccc cgacccagac taacggttct aacgttccct
     3421 tcaagccacg agggagagag ttttcctttg aggcctggaa tgccaaaatc acggacctaa
     3481 aacagaaagt tgaaaatctc ttcaatgaga aatgtgggga agctcttggc cttaaacaag
     3541 ctgtgaaggt gccgttcgcg ttatttgagt ctttcccgga agacttttat gtggaaggct
     3601 tacctgaggg tgtgccattc cgaagaccat cgacttttgg cattccgagg ctggagaaga
     3661 tactcagaaa caaagccaaa attaagttca tcattaaaaa gcccgaaatg tttgagacgg
     3721 cgattaagga gagcacctcc tctaagagcc ctcccagaaa aataaattca tcacccaatg
     3781 ttaatactac tgcatcaggt gttgaagacc ttaacatcat tcaggtgaca attccagatg
     3841 atgataatga aagactctcg aaagttgaaa aagctagaca gctaagagaa caagtgaatg
     3901 acctctttag tcggaaattt ggtgaagcta ttggtatggg ttttcctgtg aaagttccct
     3961 acaggaaaat cacaattaac cctggctgtg tggtagttga tggcatgccc ccgggggtgt
     4021 ccttcaaagc ccccagctac ctggaaatca gctccatgag aaggatctta gactctgccg
     4081 agtttatcaa attcacggtc attagaccat ttccaggact tgtgattaat aaccagctgg
     4141 ttgatcagag tgagtcagaa ggccccgtga tacaagaatc agctgaacca agccagttgg
     4201 aagttccagc cacagaagaa ataaaagaga ctgatggaag ctctcagatc aagcaagaac
     4261 cagaccccac gtggatgcgg ccgctcgagg actacaaaga ccatgacggt gattataaag
     4321 atcatgacat cgactacaag gatgacgatg acaagtagtg agggcccgat atctcgacaa
     4381 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4441 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4501 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4561 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     4621 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     4681 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     4741 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     4801 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     4861 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     4921 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaaacggggg
     4981 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5041 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5101 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5161 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5221 cgtcggggcg gcaggccctg ccatagcaga tctgcgcagc tggggctcta gggggtatcc
     5281 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5341 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt
     5461 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5521 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5581 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5641 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5701 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5821 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5881 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5941 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     6001 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     6061 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     6121 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6181 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6241 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6301 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6361 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6421 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6481 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6541 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6601 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     6661 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6721 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6781 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6841 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6901 gacgagttct tctgagcggg actctggggt tcgcgaaatg accgaccaag cgacgcccaa
     6961 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     7021 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     7081 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7141 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7201 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     7261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     7861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8341 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8401 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8461 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8521 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8581 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8641 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8701 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8761 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8821 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8881 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8941 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9001 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9061 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9121 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9181 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9241 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9301 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9361 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9421 cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg ctctgatgcc
     9481 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9541 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9601 agg
//
