LOCUS       V047481                 7550 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047481
VERSION     V047481
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             890..2299
                     /label="MMP12(NM_002426)"
                     /note="MMP12(NM_002426)"
                     /gene="MMP12"
     CDS             2336..3052
                     /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             3059..3076
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3117..3341
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3387..3815
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3829..4158
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4225..5019
                     /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    5193..5314
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5363..5379)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5387..5403
                     /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(5411..5441)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5456..5477
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5765..6350)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6521..7381)
                     /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(7382..7486)
                     /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 cttgccacca tgaagtttct
      901 tctaatactg ctcctgcagg ccactgcttc tggagctctt cccctgaaca gctctacaag
      961 cctggaaaaa aataatgtgc tatttggtga aagatactta gaaaaatttt atggccttga
     1021 gataaacaaa cttccagtga caaaaatgaa atatagtgga aacttaatga aggaaaaaat
     1081 ccaagaaatg cagcacttct tgggtctgaa agtgaccggg caactggaca catctaccct
     1141 ggagatgatg cacgcacctc gatgtggagt ccccgatgtc catcatttca gggaaatgcc
     1201 aggggggccc gtatggagga aacattatat cacctacaga atcaataatt acacacctga
     1261 catgaaccgt gaggatgttg actacgcaat ccggaaagct ttccaagtat ggagtaatgt
     1321 tacccccttg aaattcagca agattaacac aggcatggct gacattttgg tggtttttgc
     1381 ccgtggagct catggagact tccatgcttt tgatggcaaa ggtggaatcc tagcccatgc
     1441 ttttggacct ggatctggca ttggagggga tgcacatttc gatgaggacg aattctggac
     1501 tacacattca ggaggcacaa acttgttcct cactgctgtt cacgagattg gccattcctt
     1561 aggtcttggc cattctagtg atccaaaggc cgtaatgttc cccacctaca aatatgttga
     1621 catcaacaca tttcgcctct ctgctgatga catacgtggc attcagtccc tgtatggaga
     1681 cccaaaagag aaccaacgct tgccaaatcc tgacaattca gaaccagctc tctgtgaccc
     1741 caatttgagt tttgatgctg tcactaccgt gggaaataag atctttttct tcaaagacag
     1801 gttcttctgg ctgaaggttt ctgagagacc aaagaccagt gttaatttaa tttcttcctt
     1861 atggccaacc ttgccatctg gcattgaagc tgcttatgaa attgaagcca gaaatcaagt
     1921 ttttcttttt aaagatgaca aatactggtt aattagcaat ttaagaccag agccaaatta
     1981 tcccaagagc atacattctt ttggttttcc taactttgtg aaaaaaattg atgcagctgt
     2041 ttttaaccca cgtttttata ggacctactt ctttgtagat aaccagtatt ggaggtatga
     2101 tgaaaggaga cagatgatgg accctggtta tcccaaactg attaccaaga acttccaagg
     2161 aatcgggcct aaaattgatg cagtcttcta ctctaaaaac aaatactact atttcttcca
     2221 aggatctaac caatttgaat atgacttcct actccaacgt atcaccaaaa cactgaaaag
     2281 caatagctgg tttggttgtg gtggaggcgg atcaggaggc ggtggatctg gatccatggt
     2341 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2401 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2461 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2521 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2581 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2641 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2701 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2761 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2821 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2881 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2941 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3001 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agctcgagca
     3061 ccatcaccat caccattagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3301 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3361 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3421 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3481 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3541 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3601 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3661 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3721 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3781 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3841 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3901 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4021 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4081 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4141 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4201 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4261 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4321 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4381 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4441 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4501 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4561 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4621 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4681 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4741 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4801 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4861 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4921 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4981 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6301 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6361 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6421 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6481 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6541 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6601 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6661 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6721 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6781 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6841 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6901 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6961 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7021 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7081 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7141 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7201 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7261 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7321 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7381 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7441 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7501 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
