LOCUS       V037081                 8305 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037081
VERSION     V037081
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             927..3110
                     /label="HGF(NM_000601)"
                     /note="HGF(NM_000601)"
                     /gene="HGF"
     CDS             3148..3867
                     /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"
     polyA_signal    3905..4129
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4175..4603
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4617..4946
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5013..5807
                     /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    5981..6102
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6151..6167)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6175..6191
                     /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(6199..6229)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6244..6265
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6553..7138)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7309..8169)
                     /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(8170..8274)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaagcc actgggatcc gccaccatgt gggtgaccaa actcctgcca gccctgctgc
      961 tgcagcatgt cctcctgcat ctcctcctgc tccccatcgc catcccctat gcagagggac
     1021 aaaggaaaag aagaaataca attcatgaat tcaaaaaatc agcaaagact accctaatca
     1081 aaatagatcc agcactgaag ataaaaacca aaaaagtgaa tactgcagac caatgtgcta
     1141 atagatgtac taggaataaa ggacttccat tcacttgcaa ggcttttgtt tttgataaag
     1201 caagaaaaca atgcctctgg ttccccttca atagcatgtc aagtggagtg aaaaaagaat
     1261 ttggccatga atttgacctc tatgaaaaca aagactacat tagaaactgc atcattggta
     1321 aaggacgcag ctacaaggga acagtatcta tcactaagag tggcatcaaa tgtcagccct
     1381 ggagttccat gataccacac gaacacagct ttttgccttc gagctatcgg ggtaaagacc
     1441 tacaggaaaa ctactgtcga aatcctcgag gggaagaagg gggaccctgg tgtttcacaa
     1501 gcaatccaga ggtacgctac gaagtctgtg acattcctca gtgttcagaa gttgaatgca
     1561 tgacctgcaa tggggagagt tatcgaggtc tcatggatca tacagaatca ggcaagattt
     1621 gtcagcgctg ggatcatcag acaccacacc ggcacaaatt cttgcctgaa agatatcccg
     1681 acaagggctt tgatgataat tattgccgca atcccgatgg ccagccgagg ccatggtgct
     1741 atactcttga ccctcacacc cgctgggagt actgtgcaat taaaacatgc gctgacaata
     1801 ctatgaatga cactgatgtt cctttggaaa caactgaatg catccaaggt caaggagaag
     1861 gctacagggg cactgtcaat accatttgga atggaattcc atgtcagcgt tgggattctc
     1921 agtatcctca cgagcatgac atgactcctg aaaatttcaa gtgcaaggac ctacgagaaa
     1981 attactgccg aaatccagat gggtctgaat caccctggtg ttttaccact gatccaaaca
     2041 tccgagttgg ctactgctcc caaattccaa actgtgatat gtcacatgga caagattgtt
     2101 atcgtgggaa tggcaaaaat tatatgggca acttatccca aacaagatct ggactaacat
     2161 gttcaatgtg ggacaagaac atggaagact tacatcgtca tatcttctgg gaaccagatg
     2221 caagtaagct gaatgagaat tactgccgaa atccagatga tgatgctcat ggaccctggt
     2281 gctacacggg aaatccactc attccttggg attattgccc tatttctcgt tgtgaaggtg
     2341 ataccacacc tacaatagtc aatttagacc atcccgtaat atcttgtgcc aaaacgaaac
     2401 aattgcgagt tgtaaatggg attccaacac gaacaaacat aggatggatg gttagtttga
     2461 gatacagaaa taaacatatc tgcggaggat cattgataaa ggagagttgg gttcttactg
     2521 cacgacagtg tttcccttct cgagacttga aagattatga agcttggctt ggaattcatg
     2581 atgtccacgg aagaggagat gagaaatgca aacaggttct caatgtttcc cagctggtat
     2641 atggccctga aggatcagat ctggttttaa tgaagcttgc caggcctgct gtcctggatg
     2701 attttgttag tacgattgat ttacctaatt atggatgcac aattcctgaa aagaccagtt
     2761 gcagtgttta tggctggggc tacactggat tgatcaacta tgatggccta ttacgagtgg
     2821 cacatctcta tataatggga aatgagaaat gcagccagca tcatcgaggg aaggtgactc
     2881 tgaatgagtc tgaaatatgt gctggggctg aaaagattgg atcaggacca tgtgaggggg
     2941 attatggtgg cccacttgtt tgtgagcaac ataaaatgag aatggttctt ggtgtcattg
     3001 ttcctggtcg tggatgtgcc attccaaatc gtcctggtat ttttgtccga gtagcatatt
     3061 atgcaaaatg gatacacaaa attattttaa catataaggt accacagtca ctcgagtggc
     3121 ggtggaggaa gcggcggagg tggatctatg gtgagcaagg gcgaggagct gttcaccggg
     3181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     3301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     3361 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3601 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3661 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3721 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     3781 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3841 ctcggcatgg acgagctgta caagtagtct agagggcccg tttaaacccg ctgatcagcc
     3901 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3961 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4021 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4081 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4141 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4201 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4261 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4321 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4381 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4441 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4501 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4561 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4621 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4681 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4801 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4861 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4921 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4981 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5041 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5101 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5161 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5221 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5281 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5341 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5401 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5461 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5521 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5581 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5641 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5701 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5761 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5821 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5881 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5941 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     6001 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     6061 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     6121 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6181 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6241 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6361 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6421 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6481 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6541 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6601 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6661 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6721 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6781 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6841 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6901 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6961 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7021 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7081 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8281 tccccgaaaa gtgccacctg acgtc
//
