LOCUS       V038675                 7646 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038675
VERSION     V038675
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..3018
                     /label="DLST(NM_001933)"
                     /note="DLST(NM_001933)"
                     /gene="DLST"
     CDS             3025..3051
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3055..3081
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3085..3111
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3213..3437
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3483..3911
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3925..4254
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4321..5115
                     /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    5289..5410
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5459..5475)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5483..5499
                     /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(5507..5537)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5552..5573
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5861..6446)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6617..7477)
                     /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(7478..7582)
                     /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 tgctgtcccg atcccgctgt
     1681 gtgtctcggg cgttcagccg ctcgctctcc gccttccaga aggggaactg ccctctaggg
     1741 agacgttccc tgcctggggt ctccttatgc cagggaccag gttaccctaa cagcaggaag
     1801 gttgtcatta acaacagtgt cttcagtgtt cgctttttca gaactacagc tgtatgcaag
     1861 gatgacttgg ttacagtcaa aaccccagcg tttgcagaat ctgtcacaga gggagatgtc
     1921 aggtgggaga aagctgttgg agacacagtt gcagaagatg aagtggtttg tgagattgaa
     1981 actgacaaga catctgtgca ggttccatca ccagcaaatg gcgtgattga agctcttttg
     2041 gtacctgatg ggggaaaagt cgaaggaggc actccacttt tcacactcag gaaaactggt
     2101 gctgctcctg ctaaggccaa gccggctgaa gctcctgctg ctgcagcccc aaaagcagaa
     2161 cctacagcag cggcagttcc tccccctgca gcacccatac ccactcagat gccaccggtg
     2221 ccctcgccct cacagcctcc ttctggcaaa cctgtgtctg cagtaaaacc cactgttgcc
     2281 ccaccactag ctgagccagg agctggcaaa ggtctgcgtt cagaacatcg ggagaaaatg
     2341 aacaggatgc ggcagcgcat tgctcagcgt ctgaaggagg cccagaatac atgtgcaatg
     2401 ctgacaactt ttaatgagat tgacatgagt aacatccagg agatgagggc tcggcacaaa
     2461 gaggcttttt tgaagaaaca taacctcaaa ctaggcttca tgtcggcatt tgtgaaggcc
     2521 tcagcctttg ccttgcagga acagcctgtt gtaaatgcag tgattgacga cacaaccaaa
     2581 gaggtggtgt atagggatta tattgacatc agtgttgcag tggccacccc acggggtctg
     2641 gtggttccag tcatcaggaa tgtggaagct atgaattttg cagatattga acggaccatc
     2701 actgaactgg gagagaaggc ccgaaagaat gaacttgcca ttgaagatat ggatggcggt
     2761 accttcacca ttagcaatgg aggcgttttt ggctcgctct ttggaacacc cattatcaac
     2821 ccccctcagt ctgccatcct ggggatgcat ggcatctttg acaggccagt ggctatagga
     2881 ggcaaggtag aggtgcggcc catgatgtac gtggcactga cctatgatca ccggctgatt
     2941 gatggcagag aggctgtgac tttcctccgc aaaatcaagg cagcggtaga ggatcccaga
     3001 gtcctcctcc tggatcttgg tacctaccca tacgacgtac ctgattatgc aacttatcct
     3061 tatgatgtac ctgattatgc tagctatcct tatgatgtcc cagactacgc ataggatcca
     3121 ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc tcgagtctag
     3181 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3241 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3301 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3361 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     3421 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     3481 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3901 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     4261 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     4321 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4381 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4441 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4501 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4561 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4621 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4681 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4741 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4801 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4861 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4921 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4981 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5041 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5101 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     5161 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5221 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5281 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5341 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5401 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5461 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5521 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5581 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5641 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5701 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5761 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5821 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5881 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5941 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6001 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6061 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6121 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6181 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6241 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6301 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6361 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     6421 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6481 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6541 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6661 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6721 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6781 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6841 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6901 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6961 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7021 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7081 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7141 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7201 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7261 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7321 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7381 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7441 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7501 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7561 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7621 cgggagatct cccgatcccc tatggt
//
