LOCUS       V042537                 8041 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042537
VERSION     V042537
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..2781
                     /label="ATP6V1A(NM_001690)"
                     /note="ATP6V1A(NM_001690)"
                     /gene="ATP6V1A"
     CDS             2818..3534
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3544..3561
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3608..3832
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3878..4306
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4320..4649
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4716..5510
                     /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    5684..5805
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5854..5870)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5878..5894
                     /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(5902..5932)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5947..5968
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6256..6841)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7012..7872)
                     /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(7873..7977)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggattttt ccaagctacc caaaatactc
      961 gatgaagata aagaaagcac atttggttat gtgcatgggg tctcaggacc tgtggttaca
     1021 gcctgtgaca tggcgggtgc agccatgtat gagctggtga gagtgggcca cagcgaattg
     1081 gttggagaga ttattcgatt ggagggtgac atggctacta ttcaggtgta tgaagaaact
     1141 tctggtgtgt ctgttggaga tcctgtactt cgcactggta aacccctctc tgtagagctt
     1201 ggtcctggca ttatgggagc catttttgat ggtattcaaa gacctttgtc ggatatcagc
     1261 agtcagaccc aaagcatcta catccccaga ggagtaaacg tgtctgctct tagcagagat
     1321 atcaaatggg actttacacc ttgcaaaaac ctacgggttg gtagtcatat cactggcgga
     1381 gacatttatg gaattgtcag tgagaactcg cttatcaaac acaaaatcat gttaccccca
     1441 cgaaacagag gaactgtaac ttacattgct ccacctggga attatgatac ctctgatgtt
     1501 gtcttggagc ttgaatttga aggtgtaaag gagaagttca ccatggtgca agtatggcct
     1561 gtacgtcaag ttcgacctgt cactgagaag ctgccagcca atcatcctct gttgactggc
     1621 cagagagtcc ttgatgccct ttttccgtgt gtccagggag gaactactgc tatccctgga
     1681 gcctttggct gtggaaagac agtgatatca cagtctctat ccaagtattc taacagtgat
     1741 gtaatcatct atgtaggatg tggtgaaaga ggaaatgaga tgtctgaagt cctccgggac
     1801 ttcccagagc tcacaatgga ggttgatggt aaggtagagt caattatgaa gaggacagct
     1861 ttggtagcca atacctccaa tatgcctgtt gctgctagag aagcctctat ttatactgga
     1921 atcacactgt cagagtactt ccgtgacatg ggctatcatg tcagtatgat ggctgactct
     1981 acctctagat gggctgaggc ccttagagaa atctctggtc gtttagctga aatgcctgca
     2041 gatagtggat atccagccta tcttggtgcc cgtctggcct cgttttatga acgagcaggc
     2101 agggtgaaat gtcttggaaa tcctgaaaga gaagggagtg tcagcattgt aggagcagtt
     2161 tctccacctg gtggtgattt ttctgatcca gttacatctg ccactcttgg tatcgttcag
     2221 gtgttctggg gcttagataa gaaactagct caacgtaagc atttcccctc tgtcaattgg
     2281 ctcatcagct acagcaagta tatgcgtgcc ttggatgaat actatgacaa acacttcaca
     2341 gagttcgttc ctctgaggac gaaagctaag gaaattctgc aggaagaaga agacctggca
     2401 gaaattgtac agcttgtggg aaaggcttct ttggcagaaa cagataaaat cactctggag
     2461 gtagcaaaac ttatcaaaga tgatttccta caacaaaatg gatatactcc ttatgacagg
     2521 ttctgcccat tctacaagac agtagggatg ctgtccaaca tgattgcatt ttatgatatg
     2581 gctcgtagag ctgttgaaac cactgcccag agtgacaata aaatcacatg gtccattatt
     2641 cgtgagcaca tgggagacat cctctataaa ctttcctcca tgaaattcaa ggatccactg
     2701 aaagatggtg aggcaaagat caaaagcgac tatgcacaac ttcttgaaga catgcagaat
     2761 gcattccgta gccttgaaga tgatatcggc ggtggaggaa gcggcggagg tggatctatg
     2821 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2881 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2941 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3001 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     3061 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3121 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3181 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3241 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     3301 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3361 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3421 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3481 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggcggcc
     3541 gcacatcacc atcatcacca ttgactcgag tctagagggc ccgtttaaac ccgctgatca
     3601 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3661 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3721 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3781 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3841 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3901 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3961 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4021 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4081 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4141 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4201 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4261 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4321 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4381 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4441 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4501 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4561 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4621 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4681 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4741 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4801 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4861 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4921 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4981 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5041 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5101 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5161 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5221 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5281 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5341 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5401 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5461 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5521 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5581 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5641 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5701 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5761 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5821 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6781 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7981 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     8041 t
//
