LOCUS       V030117                 8470 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030117
VERSION     V030117
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..3960
                     /label="CPT1A(NM_001876)"
                     /note="CPT1A(NM_001876)"
                     /gene="CPT1A"
     polyA_signal    4037..4261
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4307..4735
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4749..5078
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5145..5939
                     /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    6113..6234
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6283..6299)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6307..6323
                     /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(6331..6361)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6376..6397
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6685..7270)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7441..8301)
                     /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(8302..8406)
                     /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 catggcagaa gctcaccaag ctgtggcctt tcagttcacg
     1681 gtcactccgg acgggattga cctgcggctg agccatgaag ctcttagaca aatctatctc
     1741 tctggacttc attcctggaa aaagaagttc atcagattca agaacggcat catcactggc
     1801 gtgtacccgg caagcccctc cagttggctt atcgtggtgg tgggcgtgat gacaacgatg
     1861 tacgccaaga tcgacccctc gttaggaata attgcaaaaa tcaatcggac tctggaaacg
     1921 gccaactgca tgtccagcca gacgaagaac gtggtcagcg gcgtgctgtt tggcaccggc
     1981 ctgtgggtgg ccctcatcgt caccatgcgc tactccctga aagtgctgct ctcctaccac
     2041 gggtggatgt tcactgagca cggcaagatg agtcgtgcca ccaagatctg gatgggtatg
     2101 gtcaagatct tttcaggccg aaaacccatg ttgtacagct tccagacatc gctgcctcgc
     2161 ctgccggtcc cggctgtcaa agacactgtg aacaggtatc tacagtcggt gaggcctctt
     2221 atgaaggaag aagacttcaa acggatgaca gcacttgctc aagattttgc tgtcggtctt
     2281 ggaccaagat tacagtggta tttgaagtta aaatcctggt gggctacaaa ttacgtgagc
     2341 gactggtggg aggagtacat ctacctccga ggacgagggc cgctcatggt gaacagcaac
     2401 tattatgcca tggatctgct gtatatcctt ccaactcaca ttcaggcagc aagagccggc
     2461 aacgccatcc atgccatcct gctttacagg cgcaaactgg accgggagga aatcaaacca
     2521 attcgtcttt tgggatccac gattccactc tgctccgctc agtgggagcg gatgtttaat
     2581 acttcccgga tcccaggaga ggagacagac accatccagc acatgagaga cagcaagcac
     2641 atcgtcgtgt accatcgagg acgctacttc aaggtctggc tctaccatga tgggcggctg
     2701 ctgaagcccc gggagatgga gcagcagatg cagaggatcc tggacaatac ctcggagcct
     2761 cagcccgggg aggccaggct ggcagccctc accgcaggag acagagttcc ctgggccagg
     2821 tgtcgtcagg cctattttgg acgtgggaaa aataagcagt ctcttgatgc tgtggagaaa
     2881 gcagcgttct ttgtgacgtt agatgaaact gaagaaggat acagaagtga agacccggat
     2941 acgtcaatgg acagctacgc caaatctcta ctacacggcc gatgttacga caggtggttt
     3001 gacaagtcgt tcacgtttgt tgtcttcaaa aacgggaaga tgggcctcaa cgctgaacac
     3061 tcctgggcag atgcgccgat cgtggcccac ctttgggagt acgtcatgtc cattgacagc
     3121 ctccagctgg gctatgcgga ggatgggcac tgcaaaggcg acatcaatcc gaacattccg
     3181 taccccacca ggctgcagtg ggacatcccg ggggaatgtc aagaggttat agagacctcc
     3241 ctgaacaccg caaatcttct ggcaaacgac gtggatttcc attccttccc attcgtagcc
     3301 tttggtaaag gaatcatcaa gaaatgtcgc acgagcccag acgcctttgt gcagctggcc
     3361 ctccagctgg cgcactacaa ggacatgggc aagttttgcc tcacatacga ggcctccatg
     3421 acccggctct tccgagaggg gaggacggag accgtgcgct cctgcaccac tgagtcatgc
     3481 gacttcgtgc gggccatggt ggacccggcc cagacggtgg aacagaggct gaagttgttc
     3541 aagttggcgt ctgagaagca tcagcatatg tatcgcctcg ccatgaccgg ctctgggatc
     3601 gatcgtcacc tcttctgcct ttacgtggtg tctaaatatc tcgctgtgga gtcccctttc
     3661 cttaaggaag ttttatctga gccttggaga ttatcaacaa gccagacccc tcagcagcaa
     3721 gtggagctgt ttgacttgga gaataaccca gagtacgtgt ccagcggagg gggctttgga
     3781 ccggttgctg atgacggcta tggtgtgtcg tacatccttg tgggagagaa cctcatcaat
     3841 ttccacattt cttccaagtt ctcttgccct gagacggatt ctcatcgctt tggaaggcac
     3901 ctgaaagaag caatgactga catcatcact ttgtttggtc tcagttctaa ttccaaaaag
     3961 taaggatcct gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     4021 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4081 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4141 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4201 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4261 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4321 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4381 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4441 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4501 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4561 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4621 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4681 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4741 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4801 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4861 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4921 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4981 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5041 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5101 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5161 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5221 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5281 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5341 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5401 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5461 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5521 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5581 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5641 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5701 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5761 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5821 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5881 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5941 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6001 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6061 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6121 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6181 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6241 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6301 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6361 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7201 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8461 cccctatggt
//
