LOCUS       V032749                 7489 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032749
VERSION     V032749
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..2979
                     /label="DRD1(NM_000794)"
                     /note="DRD1(NM_000794)"
                     /gene="DRD1"
     polyA_signal    3056..3280
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3326..3754
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3768..4097
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4164..4958
                     /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    5132..5253
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5302..5318)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5326..5342
                     /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(5350..5380)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5395..5416
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5704..6289)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6460..7320)
                     /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(7321..7425)
                     /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 catgaggact ctgaacacct ctgccatgga cgggactggg
     1681 ctggtggtgg agagggactt ctctgttcgt atcctcactg cctgtttcct gtcgctgctc
     1741 atcctgtcca cgctcctggg gaacacgctg gtctgtgctg ccgttatcag gttccgacac
     1801 ctgcggtcca aggtgaccaa cttctttgtc atctccttgg ctgtgtcaga tctcttggtg
     1861 gccgtcctgg tcatgccctg gaaggcagtg gctgagattg ctggcttctg gccctttggg
     1921 tccttctgta acatctgggt ggcctttgac atcatgtgct ccactgcatc catcctcaac
     1981 ctctgtgtga tcagcgtgga caggtattgg gctatctcca gccctttccg gtatgagaga
     2041 aagatgaccc ccaaggcagc cttcatcctg atcagtgtgg catggacctt gtctgtactc
     2101 atctccttca tcccagtgca gctcagctgg cacaaggcaa aacccacaag cccctctgat
     2161 ggaaatgcca cttccctggc tgagaccata gacaactgtg actccagcct cagcaggaca
     2221 tatgccatct catcctctgt aataagcttt tacatccctg tggccatcat gattgtcacc
     2281 tacaccagga tctacaggat tgctcagaaa caaatacggc gcattgcggc cttggagagg
     2341 gcagcagtcc acgccaagaa ttgccagacc accacaggta atggaaagcc tgtcgaatgt
     2401 tctcaaccgg aaagttcttt taagatgtcc ttcaaaagag aaactaaagt cctgaagact
     2461 ctgtcggtga tcatgggtgt gtttgtgtgc tgttggctac ctttcttcat cttgaactgc
     2521 attttgccct tctgtgggtc tggggagacg cagcccttct gcattgattc caacaccttt
     2581 gacgtgtttg tgtggtttgg gtgggctaat tcatccttga accccatcat ttatgccttt
     2641 aatgctgatt ttcggaaggc attttcaacc ctcttaggat gctacagact ttgccctgcg
     2701 acgaataatg ccatagagac ggtgagtatc aataacaatg gggccgcgat gttttccagc
     2761 catcatgagc cacgaggctc catctccaag gagtgcaatc tggtttacct gatcccacat
     2821 gctgtgggct cctctgagga cctgaaaaag gaggaggcag ctggcatcgc cagacccttg
     2881 gagaagctgt ccccagccct atcagtcata ttggactatg acactgacgt ctctctggag
     2941 aagatccaac ccatcacaca aaacggtcag cacccaacct gaggatcctg atatccagca
     3001 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3061 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3121 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3181 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3241 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3301 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3721 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4021 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4081 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4141 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4201 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4261 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4321 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4381 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4441 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4501 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4561 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4621 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4681 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4741 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4801 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4861 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4921 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4981 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5041 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5101 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5161 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5221 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5281 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7441 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
