LOCUS       V021005                 7366 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021005
VERSION     V021005
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             1660..2853
                     /label="DAP3(NM_001199849)"
                     /note="DAP3(NM_001199849)"
                     /gene="DAP3"
     polyA_signal    2933..3157
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3203..3631
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3645..3974
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4041..4835
                     /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    5009..5130
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5179..5195)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5203..5219
                     /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(5227..5257)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5272..5293
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5581..6166)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6337..7197)
                     /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(7198..7302)
                     /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 tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tgatgctgaa aggaataaca
     1681 aggcttatct ctaggatcca taagttggac cctgggcgtt ttttacacat ggggacccag
     1741 gctcgccaaa gcattgctgc tcacctagat aaccaggttc cagttgagag tccgagagct
     1801 atttcccgca ccaatgagaa tgacccggcc aagcatgggg atcagcacga gggtcagcac
     1861 tacaacatct ccccccagga tttggagact gtatttcccc atggccttcc tcctcgcttt
     1921 gtgatgcagg tgaagacatt cagtgaagct tgcctgatgg taaggaaacc agccctagaa
     1981 cttctgcatt acctgaaaaa caccagtttt gcttatccag ctatacgata tcttctgtat
     2041 ggagagaagg gaacaggaaa aaccctaagt ctttgccatg ttattcattt ctgtgcaaaa
     2101 caggactggc tgatactaca tattccagat gctcatcttt gggtgaaaaa ttgtcgggat
     2161 cttctgcagt ccagctacaa caaacagcgc tttgatcaac ctttagaggc ttcaacctgg
     2221 ctgaagaatt tcaaaactac aaatgagcgc ttcctgaacc agataaaagt tcaagagaag
     2281 tatgtctgga ataagagaga aagcactgag aaagggagtc ctctgggaga agtggttgaa
     2341 cagggcataa cacgggtgag gaacgccaca gatgcagttg gaattgtgct gaaagagcta
     2401 aagaggcaaa gttctttggg tatgtttcac ctcctagtgg ccgtggatgg aatcaatgct
     2461 ctttggggaa gaaccactct gaaaagagaa gataaaagcc cgattgcccc cgaggaatta
     2521 gcacttgttc acaacttgag gaaaatgatg aaaaatgatt ggcatggagg cgccattgtg
     2581 tcggctttga gccagactgg gtctctcttt aagccccgga aagcctatct gccccaggag
     2641 ttgctgggaa aggaaggatt tgatgccctg gatcccttta ttcccatcct ggtttccaac
     2701 tataacccaa aggaatttga aagttgtatt cagtattatt tggaaaacaa ttggcttcaa
     2761 catgagaaag ctcctacaga agaagggaaa aaagagctgc tgttcctaag taacgcgaac
     2821 ccctcgctgc tggagcggca ctgtgcctac ctctaagaat tctgcagata tccagcacag
     2881 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2941 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3001 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3061 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3121 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3181 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3541 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3601 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3661 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3721 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3841 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3901 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3961 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4021 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4081 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4141 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4201 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4261 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4321 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4381 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4441 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4501 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4561 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4621 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4681 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4741 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4801 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4861 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4921 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4981 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5161 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5221 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5281 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5341 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5401 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5461 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5521 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5581 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5641 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5701 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5761 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5821 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5881 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5941 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6001 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6061 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6241 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6301 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6361 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6421 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6481 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6541 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6601 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6661 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6721 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6781 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6841 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6901 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6961 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7021 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7081 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7141 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7201 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7261 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7321 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
