LOCUS       V033512                 9920 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033512
VERSION     V033512
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             908..1624
                     /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             1667..5383
                     /label="POLG(NM_002693)"
                     /note="POLG(NM_002693)"
                     /gene="POLG"
     CDS             5390..5413
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             5420..5446
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    5487..5711
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5757..6185
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6199..6528
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6595..7389
                     /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    7563..7684
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7733..7749)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7757..7773
                     /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(7781..7811)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7826..7847
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8135..8720)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8891..9751)
                     /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(9752..9856)
                     /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 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1081 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1321 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1381 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1501 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1621 caagggcggt ggaggcagtg gtggaggcgg tagcgaattc gccaccatga gccgcctgct
     1681 ctggaggaag gtggccggcg ccaccgtcgg gccagggccg gttccagctc cggggcgctg
     1741 ggtctccagc tccgtccccg cgtccgaccc cagcgacggg cagcggcggc ggcagcagca
     1801 gcagcagcag cagcagcagc agcaacagca gcctcagcag ccgcaagtgc tatcctcgga
     1861 gggcgggcag ctgcggcaca acccattgga catccagatg ctctcgagag ggctgcacga
     1921 gcaaatcttc gggcaaggag gggagatgcc tggcgaggcc gcggtgcgcc gcagcgtcga
     1981 gcacctgcag aagcacgggc tctgggggca gccagccgtg cccttgcccg acgtggagct
     2041 gcgcctgccg cccctctacg gggacaacct ggaccagcac ttccgcctcc tggcccagaa
     2101 gcagagcctg ccctacctgg aggcggccaa cttgctgttg caggcccagc tgcccccgaa
     2161 gcccccggct tgggcctggg cggagggctg gacccggtac ggccccgagg gggaggccgt
     2221 acccgtggcc atccccgagg agcgggccct ggtgttcgac gtggaggtct gcttggcaga
     2281 gggaacttgc cccacattgg cggtggccat atccccctcg gcctggtatt cctggtgcag
     2341 ccagcggctg gtggaagagc gttactcttg gaccagccag ctgtcgccgg ctgacctcat
     2401 ccccctggag gtccctactg gtgccagcag ccccacccag agagactggc aggagcagtt
     2461 agtggtgggg cacaatgttt cctttgaccg agctcatatc agggagcagt acctgatcca
     2521 gggttcccgc atgcgtttcc tggacaccat gagcatgcac atggccatct cagggctaag
     2581 cagcttccag cgcagtctgt ggatagcagc caagcagggc aaacacaagg tccagccccc
     2641 cacaaagcaa ggccagaagt cccagaggaa agccagaaga ggcccagcga tctcatcctg
     2701 ggactggctg gacatcagca gtgtcaacag tctggcagag gtgcacagac tttatgtagg
     2761 ggggcctccc ttagagaagg agcctcgaga actgtttgtg aagggcacca tgaaggacat
     2821 tcgtgagaac ttccaggacc tgatgcagta ctgtgcccag gacgtgtggg ccacccatga
     2881 ggttttccag cagcagctac cgctcttctt ggagaggtgt ccccacccag tgactctggc
     2941 cggcatgctg gagatgggtg tctcctacct gcctgtcaac cagaactggg agcgttacct
     3001 ggcagaggca cagggcactt atgaggagct ccagcgggag atgaagaagt cgttgatgga
     3061 tctggccaat gatgcctgcc agctgctctc aggagagagg tacaaagaag acccctggct
     3121 ctgggacctg gagtgggacc tgcaagaatt taagcagaag aaagctaaga aggtgaagaa
     3181 ggaaccagcc acagccagca agttgcccat cgagggggct ggggcccctg gtgatcccat
     3241 ggatcaggaa gacctcggcc cctgcagtga ggaggaggag tttcaacaag atgtcatggc
     3301 ccgcgcctgc ttgcagaagc tgaaggggac cacagagctc ctgcccaagc ggccccagca
     3361 ccttcctgga caccctggat ggtaccggaa gctctgcccc cggctagacg accctgcatg
     3421 gaccccgggc cccagcctcc tcagcctgca gatgcgggtc acacctaaac tcatggcact
     3481 tacctgggat ggcttccctc tgcactactc agagcgtcat ggctggggct acttggtgcc
     3541 tgggcggcgg gacaacctgg ccaagctgcc gacaggtacc accctggagt cagctggggt
     3601 ggtctgcccc tacagagcca tcgagtccct gtacaggaag cactgtctcg aacaggggaa
     3661 gcagcagctg atgccccagg aggccggcct ggcggaggag ttcctgctca ctgacaatag
     3721 tgccatatgg caaacggtag aagaactgga ttacttagaa gtggaggctg aggccaagat
     3781 ggagaacttg cgagctgcag tgccaggtca acccctagct ctgactgccc gtggtggccc
     3841 caaggacacc cagcccagct atcaccatgg caatggacct tacaacgacg tggacatccc
     3901 tggctgctgg tttttcaagc tgcctcacaa ggatggtaat agctgtaatg tgggaagccc
     3961 ctttgccaag gacttcctgc ccaagatgga ggatggcacc ctgcaggctg gcccaggagg
     4021 tgccagtggg ccccgtgctc tggaaatcaa caaaatgatt tctttctgga ggaacgccca
     4081 taaacgtatc agctcccaga tggtggtgtg gctgcccagg tcagctctgc cccgtgctgt
     4141 gatcaggcac cccgactatg atgaggaagg cctctatggg gccatcctgc cccaagtggt
     4201 gactgccggc accatcactc gccgggctgt ggagcccaca tggctcaccg ccagcaatgc
     4261 ccggcctgac cgagtaggca gtgagttgaa agccatggtg caggccccac ctggctacac
     4321 ccttgtgggt gctgatgtgg actcccaaga gctgtggatt gcagctgtgc ttggagacgc
     4381 ccactttgcc ggcatgcatg gctgcacagc ctttgggtgg atgacactgc agggcaggaa
     4441 gagcaggggc actgatctac acagtaagac agccactact gtgggcatca gccgtgagca
     4501 tgccaaaatc ttcaactacg gccgcatcta tggtgctggg cagccctttg ctgagcgctt
     4561 actaatgcag tttaaccacc ggctcacaca gcaggaggca gctgagaagg cccagcagat
     4621 gtacgctgcc accaagggcc tccgctggta tcggctgtcg gatgagggcg agtggctggt
     4681 gagggagttg aacctcccag tggacaggac tgagggtggc tggatttccc tgcaggatct
     4741 gcgcaaggtc cagagagaaa ctgcaaggaa gtcacagtgg aagaagtggg aggtggttgc
     4801 tgaacgggca tggaaggggg gcacagagtc agaaatgttc aataagcttg agagcattgc
     4861 tacgtctgac ataccacgta ccccggtgct gggctgctgc atcagccgag ccctggagcc
     4921 ctcggctgtc caggaagagt ttatgaccag ccgtgtgaat tgggtggtac agagctctgc
     4981 tgttgactac ttacacctca tgcttgtggc catgaagtgg ctgtttgaag agtttgccat
     5041 agatgggcgc ttctgcatca gcatccatga cgaggttcgc tacctggtgc gggaggagga
     5101 ccgctaccgc gctgccctgg ccttgcagat caccaacctc ttgaccaggt gcatgtttgc
     5161 ctacaagctg ggtctgaatg acttgcccca gtcagtcgcc tttttcagtg cagtcgatat
     5221 tgaccggtgc ctcaggaagg aagtgaccat ggattgtaaa accccttcca acccaactgg
     5281 gatggaaagg agatacggga ttccccaggg tgaagcgctg gatatttacc agataattga
     5341 actcaccaaa ggctccttgg aaaaacgaag ccagcctgga ccagatatcg actacaagga
     5401 cgacgatgac aagctcgagt acccatacga cgtaccagat tacgcttagt ctagagggcc
     5461 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5521 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5581 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5641 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5701 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     6001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6181 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     6481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     6541 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     6601 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     6661 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6721 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     6781 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6841 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6901 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6961 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     7021 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     7081 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     7141 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     7201 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     7261 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     7321 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     7381 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     7441 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     7501 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     7561 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7621 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7681 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7741 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7801 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7921 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7981 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     8041 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     8101 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     8161 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     8221 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8641 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     8701 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8761 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8821 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8881 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     8941 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     9001 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     9061 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     9121 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     9181 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     9241 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     9301 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     9361 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     9421 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     9481 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     9541 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     9601 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     9661 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9721 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9781 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9841 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     9901 atctcccgat cccctatggt
//
