LOCUS       V046518                 8130 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046518
VERSION     V046518
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..3621
                     /label="PABPC4(NM_001135653)"
                     /note="PABPC4(NM_001135653)"
                     /gene="PABPC4"
     polyA_signal    3697..3921
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3967..4395
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4409..4738
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4805..5599
                     /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    5773..5894
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5943..5959)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5967..5983
                     /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(5991..6021)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6036..6057
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6345..6930)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7101..7961)
                     /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(7962..8066)
                     /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 catgaacgct gcggccagca gctaccccat ggcctccctg
     1681 tacgtgggcg acctgcattc ggacgtcacc gaggccatgc tgtacgaaaa gttcagcccc
     1741 gcggggcctg tgctgtccat ccgggtctgc cgcgatatga tcacccgccg ctccctgggc
     1801 tatgcctacg tcaacttcca gcagccggcc gacgctgagc gggctttgga caccatgaac
     1861 tttgatgtga ttaagggaaa gccaatccgc atcatgtggt ctcagaggga tccctctttg
     1921 agaaaatctg gtgtgggaaa cgtcttcatc aagaacctgg acaaatctat agataacaag
     1981 gcactttatg atactttttc tgcttttgga aacatactgt cctgcaaggt ggtgtgtgat
     2041 gagaacggct ctaagggtta tgcctttgtc cacttcgaga cccaagaggc tgccgacaag
     2101 gccatcgaga agatgaatgg catgctcctc aatgaccgca aagtatttgt gggcagattc
     2161 aagtctcgca aagagcggga agctgagctt ggagccaaag ccaaggaatt caccaatgtt
     2221 tatatcaaaa actttgggga agaggtggat gatgagagtc tgaaagagct attcagtcag
     2281 tttggtaaga ccctaagtgt caaggtgatg agagatccca atgggaaatc caaaggcttt
     2341 ggctttgtga gttacgaaaa acacgaggat gccaataagg ctgtggaaga gatgaatgga
     2401 aaagaaataa gtggtaaaat catatttgta ggccgtgcac aaaagaaagt agaacggcag
     2461 gcagagttaa aacggaaatt tgaacagttg aaacaggaga gaattagtcg atatcagggg
     2521 gtgaatctct acattaagaa cttggatgac actattgatg atgagaaatt aaggaaagaa
     2581 ttttctcctt ttggatcaat taccagtgct aaggtaatgc tggaggatgg aagaagcaaa
     2641 gggtttggct tcgtctgctt ctcatctcct gaagaagcaa ccaaagcagt cactgagatg
     2701 aatggacgca ttgtgggctc caagccacta tatgttgccc tggcccagag gaaggaagag
     2761 agaaaggctc acctgaccaa ccagtatatg caacgagtgg ctggaatgag agcacttcct
     2821 gccaatgcca tcttaaatca gttccagcct gcagcgggtg gctactttgt gccagcagtc
     2881 ccacaggctc agggaaggcc tccatattat acacctaacc agttagcaca gatgaggcct
     2941 aatccacgct ggcagcaagg tgggagacct caaggcttcc aaggaatgcc aagtgctata
     3001 cgccagtctg ggcctcgtcc aactcttcgc catctggctc caactggtaa tgctccggcc
     3061 tctcgtggcc tccctactac cactcagaga gtcgggtctg agtgcccgga ccgcttggct
     3121 atggactttg gtggggctgg tgccgcccag caagggctga ctgacagctg ccagtctgga
     3181 ggcgttccca cagctgtgca gaacttagcg ccacgcgctg ctgttgctgc tgctgctccc
     3241 cgggctgttg ccccctacaa atacgcctcc agtgtccgca gccctcatcc tgccatacag
     3301 cctctgcagg caccccagcc tgcggtccat gtgcaggggc aggagccact gactgcctcc
     3361 atgctggctg cagcaccccc ccaggaacag aagcagatgc tgggagaacg cttgttccca
     3421 ctcatccaaa caatgcattc aaatctggct gggaagatca cgggaatgct gctggagata
     3481 gacaactctg agctgctgca catgttagag tcccccgagt ctctccgctc caaggtggat
     3541 gaagctgtag cagttctaca ggctcatcat gccaagaaag aagctgccca gaaggtgggc
     3601 gctgttgctg ctgctacctc ttaggatcct gatatccagc acagtggcgg ccgctcgagt
     3661 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3721 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3781 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3841 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3901 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3961 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4021 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4081 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4141 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4201 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4261 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4321 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4381 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4501 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4561 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4621 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4681 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4741 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4801 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4861 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4921 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4981 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5041 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5101 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5161 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5221 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5281 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5341 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5401 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5461 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5521 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5581 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5641 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5701 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5761 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5881 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5941 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6001 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6061 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6121 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6181 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6241 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6301 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6361 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6421 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6481 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6541 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6601 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6661 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6721 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6781 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6841 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag
     6901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8101 ggatcgggag atctcccgat cccctatggt
//
