LOCUS       V030120                 7819 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030120
VERSION     V030120
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             874..1590
                     /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             1642..3309
                     /label="TCP1(NM_030752)"
                     /note="TCP1(NM_030752)"
                     /gene="TCP1"
     polyA_signal    3386..3610
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3656..4084
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4098..4427
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4494..5288
                     /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    5462..5583
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5632..5648)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5656..5672
                     /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(5680..5710)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5725..5746
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6034..6619)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6790..7650)
                     /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(7651..7755)
                     /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggagggg cctttgtccg tgttcggtga ccgcagcact
     1681 ggggaaacga tccgctccca aaacgttatg gctgcagctt cgattgccaa tattgtaaaa
     1741 agttctcttg gtccagttgg cttggataaa atgttggtgg atgatattgg tgatgtaacc
     1801 attactaacg atggtgcaac catcctgaag ttactggagg tagaacatcc tgcagctaaa
     1861 gttctttgtg agctggctga tctgcaagac aaagaagttg gagatggaac tacttcagtg
     1921 gttattattg cagcagaact cctaaaaaat gcagatgaat tagtcaaaca gaaaattcat
     1981 cccacatcag ttattagtgg ctatcgactt gcttgcaagg aagcagtgcg ttatatcaat
     2041 gaaaacctaa ttgttaacac agatgaactg ggaagagatt gcctgattaa tgctgctaag
     2101 acatccatgt cttccaaaat cattggaata aatggtgatt tctttgctaa catggtagta
     2161 gatgctgtac ttgctattaa atacacagac ataagaggcc agccacgcta tccagtcaac
     2221 tctgttaata ttttgaaagc ccatgggaga agtcaaatgg agagtatgct catcagtggc
     2281 tatgcactca actgtgtggt gggatcccag ggcatgccca agagaatcgt aaatgcaaaa
     2341 attgcttgcc ttgacttcag cctgcaaaaa acaaaaatga agcttggtgt acaggtggtc
     2401 attacagacc ctgaaaaact ggaccaaatt agacagagag aatcagatat caccaaggag
     2461 agaattcaga agatcctggc aactggtgcc aatgttattc taaccactgg tggaattgat
     2521 gatatgtgtc tgaagtattt tgtggaggct ggtgctatgg cagttagaag agttttaaaa
     2581 agggacctta aacgcattgc caaagcttct ggagcaacta ttctgtcaac cctggccaat
     2641 ttggaaggtg aagaaacttt tgaagctgca atgttgggac aggcagaaga agtggtacag
     2701 gagagaattt gtgatgatga gctgatctta atcaaaaata ctaaggctcg tacgtctgca
     2761 tcgattatct tacgtggggc aaatgatttc atgtgtgatg agatggagcg ctctttacat
     2821 gatgcacttt gtgtagtgaa gagagttttg gagtcaaaat ctgtggttcc cggtgggggt
     2881 gctgtagaag cagccctttc catatacctt gaaaactatg caaccagcat ggggtctcgg
     2941 gaacagcttg cgattgcaga gtttgcaaga tcacttcttg ttattcccaa tacactagca
     3001 gttaatgctg cccaggactc cacagatctg gttgcaaaat taagagcttt tcataatgag
     3061 gcccaggtta acccagaacg taaaaatcta aaatggattg gtcttgattt gagcaatggt
     3121 aaacctcgag acaacaaaca agcaggggtg tttgaaccaa ccatagttaa agttaagagt
     3181 ttgaaatttg caacagaagc tgcaatcacc attcttcgaa ttgatgatct tattaaatta
     3241 catccagaaa gtaaagatga taaacatgga agttatgaag atgctgttca ctctggagcc
     3301 cttaatgatt gaggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc
     3361 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3421 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3481 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3541 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3601 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4081 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4141 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4261 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4321 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4381 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4441 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4501 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4561 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4621 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4681 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4741 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4801 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4861 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4921 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4981 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5041 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5101 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5161 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5221 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5281 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5341 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5401 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5461 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5521 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5581 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5641 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5701 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5761 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5821 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5881 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5941 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6001 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6061 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6121 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6181 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6241 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6301 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6361 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6421 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6481 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6541 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6601 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6661 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6721 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6781 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6841 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6901 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6961 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7021 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7081 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7141 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7201 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7261 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7321 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7381 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7441 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7501 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7561 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7621 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7681 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7741 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7801 tctcccgatc ccctatggt
//
