LOCUS       V041313                 8920 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041313
VERSION     V041313
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..4410
                     /label="PCDHGB4(NM_003736)"
                     /note="PCDHGB4(NM_003736)"
                     /gene="PCDHGB4"
     polyA_signal    4487..4711
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4757..5185
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5199..5528
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5595..6389
                     /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    6563..6684
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6733..6749)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6757..6773
                     /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(6781..6811)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6826..6847
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7135..7720)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7891..8751)
                     /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(8752..8856)
                     /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 catggggagc ggcgccgggg agctgggccg ggctgagagg
     1681 ctgccagtgc tctttctctt cctgctgtct ttgttctgcc cggcgctctg tgagcagatc
     1741 cgctacagga ttcccgagga aatgcccaag ggctccgtag tggggaacct cgccacggac
     1801 ctggggttca gcgtccagga gttaccgact cgaaaactgc gcgtcagttc ggagaagcct
     1861 tacttcaccg tgagcgcaga gagcggggag ttgcttgtga gcagcaggct agacagggag
     1921 gagatatgcg ggaagaagcc agcttgtgct ctggaatttg aggctgttgc tgaaaatcca
     1981 ctgaactttt atcacgtgaa tgtggagatc gaggacatta atgaccacac gccaaaattc
     2041 acgcaaaatt cctttgagct gcaaataagt gagtctgcac agcctggcac acgatttata
     2101 ttaggatctg cccatgatgc ggatattggt agcaacacac tgcagaatta ccaactcagt
     2161 cccagtgatc atttctcact gataaataaa gagaaatcag atggcagtaa ataccctgag
     2221 atggtattga agacaccttt ggacagagaa aagcagaaat cctaccactt gactttgact
     2281 gccttggact ttggagctcc acccctaagc agcactgcac agatacacgt tctagtgact
     2341 gatgccaatg ataatgctcc agtgttcagt caagacgtat acagggtgag cctttcagaa
     2401 aacgtgtacc cggggaccac ggtgctacag gtgactgcca cggaccagga tgagggtgtc
     2461 aatgccgaga ttactttctc tttcagtgaa gctagccaga tcacccaatt tgacctgaac
     2521 tctaacaccg gggaaattac tgttttaaat acattagatt ttgaagaagt caaagaatat
     2581 tccatagttt tggaagcaag ggacggtgga ggaatgattg cgcaatgcac agtggaggta
     2641 gaagtcatag atgaaaatga caacgcccca gaagtgatat tccagtctct acccaaccta
     2701 attatggagg acgccgagct gggaacacat attgctttgc tcaaagtccg tgacaaggat
     2761 tccagacaca atggagaagt gacttgtaaa ttggaaggtg atgttccatt taaaatatta
     2821 acttcttcaa gaaacacgta taaattagtg acagatgctg ttctagaccg cgagcagaat
     2881 ccagagtaca atataaccgt tacggcaaca gatcggggca agcctcccct ctcctccagt
     2941 tccagcatca ccctgcacat tggtgatgta aatgacaacg ctccggtttt ctcacagtct
     3001 tcctatatag tccacgtggc cgagaacaac ccgcctggag cctctatttc acaagtcagg
     3061 gcttctgatc cggacttggg gcccaacggc caagtctctt actgcatcat ggccagtgac
     3121 ctggagcagc gggagctgtc atcctacgtg tccataagcg cggagagcgg ggtggtgttc
     3181 gcgcagcgcg ccttcgacca cgagcagctg cgcgccttcg aactcacact gcaggcccgc
     3241 gaccagggct cgccagcgct cagcgcgaac gtgagcctgc gcgtgttagt ggacgaccgc
     3301 aacgacaatg cgccacgggt gctgtacccc gcgctgggtc ccgacggctc tgcgctcttc
     3361 gatatggtgc cgcacgctgc agagcctggc tacttggtga ccaaggtagt ggcggtggac
     3421 gcagactcag gacacaacgc ctggctgtcc taccacgtgc tgcaggctag cgagcccggg
     3481 ctcttcagcc tggggctgcg cacgggcgaa gtgcgcacag cgcgtgcctt aggcgacagg
     3541 gacgccgtcc gccagcgcct tctggtcgcc gtgcgtgacg gtggacagcc accactctcg
     3601 gccactgcca cgttgcacct ggtcttcgcc gacagcttgc aggaggtgct gccggatatc
     3661 actgaccgcc ccgacccctc tgacctccag gctgagctgc agttttacct agtggtggcc
     3721 ttggccttga tctcagtgct cttcctcgtg gccatgattc tggccattgc cttgcgcctg
     3781 cgacgctcct ccagccccgc ctcctggagc tgcttccagc ctggtctctg tgttaaatcc
     3841 gaatccgtgg ttccccccaa ctacagcgag gggactttgc cttattccta caatctatgt
     3901 gttgcacata caggaaagac ggagtttaat ttcctaaaat gtagtgagca gttgagttca
     3961 ggacaagaca tactttgcgg tgattcatct ggggccttat ttccactttg taattccagt
     4021 gaattgactt cccatcagca agccccgccc aacacggact ggcgtttctc tcaggcccag
     4081 agacccggca ccagcggctc ccaaaatggc gatgacaccg gcacctggcc caacaaccag
     4141 tttgacacag agatgctgca agccatgatc ttggcgtccg ccagtgaagc tgctgatggg
     4201 agctccaccc tgggaggggg tgccggcacc atgggattga gcgcccgcta cggaccccag
     4261 ttcaccctgc agcacgtgcc cgactaccgc cagaatgtct acatcccagg cagcaatgcc
     4321 acactgacca acgcagctgg caagcgggat ggcaaggccc cagcaggtgg caatggcaac
     4381 aagaagaagt cgggcaagaa ggagaagaag taaggatcct gatatccagc acagtggcgg
     4441 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     4501 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4561 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4621 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4681 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     4741 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4801 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4861 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4921 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4981 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5041 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5101 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5161 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5221 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5281 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5341 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5401 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5461 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5521 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5581 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5641 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5701 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5761 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5821 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5881 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5941 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6001 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6061 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6121 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6181 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6241 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6301 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6361 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     6421 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     6481 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6541 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6601 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6661 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6721 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6781 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6841 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7561 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     7681 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7741 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7801 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7861 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7921 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7981 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8041 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8101 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8161 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8221 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8281 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8341 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     8401 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     8461 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     8521 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     8581 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8641 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8701 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8761 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8821 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8881 tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
