LOCUS       V036953                 8839 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036953
VERSION     V036953
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..4329
                     /label="EPS15(NM_001981)"
                     /note="EPS15(NM_001981)"
                     /gene="EPS15"
     polyA_signal    4406..4630
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4676..5104
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5118..5447
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5514..6308
                     /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    6482..6603
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6652..6668)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6676..6692
                     /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(6700..6730)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6745..6766
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7054..7639)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7810..8670)
                     /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(8671..8775)
                     /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 catggctgcg gcggcccagc tctctctgac acagttatca
     1681 agtgggaatc ctgtatatga aaaatactat agacaggttg atacaggcaa tactggaagg
     1741 gtgttggctt ctgatgctgc tgctttcctg aaaaaatcag ggcttccaga cttgatactt
     1801 ggaaagattt gggatttagc cgacacagat ggcaaaggta tcctgaacaa acaagaattc
     1861 tttgttgctt tgcgtcttgt ggcatgtgcc cagaatggat tggaagtttc actaagtagt
     1921 ttgaacctgg ctgttcctcc accaagattt catgatacca gtagtccttt gctaatcagt
     1981 ggaacctctg cagctgagct cccatgggct gtaaaacctg aagataaggc caaatatgat
     2041 gcaatatttg atagtttaag cccagtgaat ggatttctgt ctggtgataa agtgaaacca
     2101 gtgttgctca actctaagtt acctgtggat atccttggaa gagtttggga gttgagtgat
     2161 attgaccatg atggaatgct tgacagagat gagtttgcag ttgccatgtt tttggtatac
     2221 tgtgcactgg agaaagaacc tgtgccaatg tccttgcctc cagccttggt gccaccatct
     2281 aagagaaaaa cgtgggttgt atcccctgca gaaaaagcta aatatgatga aatcttcctg
     2341 aaaactgata aagatatgga cggatttgtg tctggattgg aggtccgtga aatattcttg
     2401 aaaacaggtt taccttctac cttactagcc catatatggt cattatgcga cacaaaggac
     2461 tgtgggaagc tttcaaagga tcagtttgcc ttggcttttc acttaatcag tcagaagtta
     2521 atcaagggca ttgatcctcc tcacgttctt actcctgaaa tgattccacc atcagacagg
     2581 gccagtttac aaaagaacat cataggatca agtcctgttg cagatttctc tgctattaag
     2641 gaactagata ctcttaacaa tgaaatagtt gacctacaga gggaaaagaa taatgtggaa
     2701 caggacctta aggagaagga agatactatt aaacagagga caagtgaggt tcaggatctt
     2761 caagatgaag ttcaaaggga gaatactaat ctgcaaaaac tacaggccca gaaacagcag
     2821 gtacaggaac tccttgatga actggatgag cagaaagccc agctggagga gcaactcaag
     2881 gaagtcagaa agaaatgtgc tgaggaggcc caactgatct cttctctgaa agctgaatta
     2941 actagtcagg aatcgcagat ctccacttac gaagaagaat tggcaaaagc tagagaagag
     3001 ctgagccgtc tacagcaaga aacagcagaa ttggaggaga gtgtagagtc agggaaggct
     3061 cagttggaac ctcttcagca gcacctacaa gattcacaac aggaaattag ttcaatgcaa
     3121 atgaaactga tggaaatgaa agatttggaa aatcataata gtcagttaaa ttggtgcagt
     3181 agcccacaca gcattcttgt aaacggagct acagattatt gcagcctcag caccagcagc
     3241 agtgaaacag ccaaccttaa tgaacatgtt gaaggccaga gcaacctaga gtctgagccc
     3301 atacaccagg aatctccagc aagaagtagt cctgaactac tgccttctgg tgtgactgat
     3361 gaaaatgagg tgactacagc tgttactgaa aaagtttgtt ctgaactcga caataataga
     3421 cattcaaaag aggaagatcc atttaatgta gactcaagtt cgctgacagg tccagttgca
     3481 gatacaaact tggatttttt ccagtctgat ccttttgttg gcagtgatcc tttcaaggat
     3541 gatccttttg gaaaaatcga tccatttggt ggtgatcctt tcaaaggttc agatccattt
     3601 gcatcagact gtttcttcag gcaatctact gatccttttg ccacttcaag cactgaccct
     3661 ttcagtgcag ccaacaatag cagtattaca tcggtagaaa cgttgaagca caatgatcct
     3721 tttgctcctg gtggaacagt tgttgcagca agcgattcag ccacagaccc ctttgcttct
     3781 gtttttggga atgaatcatt tggaggtgga tttgctgact tcagcacatt gtcaaaggtc
     3841 aacaatgaag atccttttcg ttcagccaca tcgagctctg tcagcaacgt agtgattaca
     3901 aaaaatgtat ttgaggaaac atcggtcaaa agtgaagatg aacccccagc actgccacca
     3961 aagatcggaa ctccaacaag accctgccct ctaccacctg ggaaaagatc catcaacaaa
     4021 ttggattctc ctgatccctt taaactgaat gatccatttc agcctttccc aggcaacgat
     4081 agccccaaag aaaaagatcc tgaaatattt tgtgatccat tcacttctgc tactaccact
     4141 accaataaag aggctgatcc aagcaatttt gccaacttca gtgcttatcc ctctgaagaa
     4201 gatatgatcg aatgggccaa gagggaaagt gagagagagg aagagcagag gcttgcccga
     4261 ctaaatcagc aggaacaaga agacttagaa ctggctattg cactcagcaa atctgagata
     4321 tcagaagcat gaggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc
     4381 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4441 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4501 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4561 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4621 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4681 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4741 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4801 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4861 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4921 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4981 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5041 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5101 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5221 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5281 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5341 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5401 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5461 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5521 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5581 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5641 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5701 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5761 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5821 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5881 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5941 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6001 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6061 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6121 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6181 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6241 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6301 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     6361 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6421 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6481 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6601 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6661 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6721 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6781 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7561 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     7621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8821 tctcccgatc ccctatggt
//
