LOCUS       V026236                 8419 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026236
VERSION     V026236
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..3909
                     /label="MLH1(NM_000249)"
                     /note="MLH1(NM_000249)"
                     /gene="MLH1"
     polyA_signal    3986..4210
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4256..4684
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4698..5027
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5094..5888
                     /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    6062..6183
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6232..6248)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6256..6272
                     /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(6280..6310)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6325..6346
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6634..7219)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7390..8250)
                     /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(8251..8355)
                     /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 catgtcgttc gtggcagggg ttattcggcg gctggacgag
     1681 acagtggtga accgcatcgc ggcgggggaa gttatccagc ggccagctaa tgctatcaaa
     1741 gagatgattg agaactgttt agatgcaaaa tccacaagta ttcaagtgat tgttaaagag
     1801 ggaggcctga agttgattca gatccaagac aatggcaccg ggatcaggaa agaagatctg
     1861 gatattgtat gtgaaaggtt cactactagt aaactgcagt cctttgagga tttagccagt
     1921 atttctacct atggctttcg aggtgaggct ttggccagca taagccatgt ggctcatgtt
     1981 actattacaa cgaaaacagc tgatggaaag tgtgcataca gagcaagtta ctcagatgga
     2041 aaactgaaag cccctcctaa accatgtgct ggcaatcaag ggacccagat cacggtggag
     2101 gacctttttt acaacatagc cacgaggaga aaagctttaa aaaatccaag tgaagaatat
     2161 gggaaaattt tggaagttgt tggcaggtat tcagtacaca atgcaggcat tagtttctca
     2221 gttaaaaaac aaggagagac agtagctgat gttaggacac tacccaatgc ctcaaccgtg
     2281 gacaatattc gctccatctt tggaaatgct gttagtcgag aactgataga aattggatgt
     2341 gaggataaaa ccctagcctt caaaatgaat ggttacatat ccaatgcaaa ctactcagtg
     2401 aagaagtgca tcttcttact cttcatcaac catcgtctgg tagaatcaac ttccttgaga
     2461 aaagccatag aaacagtgta tgcagcctat ttgcccaaaa acacacaccc attcctgtac
     2521 ctcagtttag aaatcagtcc ccagaatgtg gatgttaatg tgcaccccac aaagcatgaa
     2581 gttcacttcc tgcacgagga gagcatcctg gagcgggtgc agcagcacat cgagagcaag
     2641 ctcctgggct ccaattcctc caggatgtac ttcacccaga ctttgctacc aggacttgct
     2701 ggcccctctg gggagatggt taaatccaca acaagtctga cctcgtcttc tacttctgga
     2761 agtagtgata aggtctatgc ccaccagatg gttcgtacag attcccggga acagaagctt
     2821 gatgcatttc tgcagcctct gagcaaaccc ctgtccagtc agccccaggc cattgtcaca
     2881 gaggataaga cagatatttc tagtggcagg gctaggcagc aagatgagga gatgcttgaa
     2941 ctcccagccc ctgctgaagt ggctgccaaa aatcagagct tggaggggga tacaacaaag
     3001 gggacttcag aaatgtcaga gaagagagga cctacttcca gcaaccccag aaagagacat
     3061 cgggaagatt ctgatgtgga aatggtggaa gatgattccc gaaaggaaat gactgcagct
     3121 tgtacccccc ggagaaggat cattaacctc actagtgttt tgagtctcca ggaagaaatt
     3181 aatgagcagg gacatgaggt tctccgggag atgttgcata accactcctt cgtgggctgt
     3241 gtgaatcctc agtgggcctt ggcacagcat caaaccaagt tataccttct caacaccacc
     3301 aagcttagtg aagaactgtt ctaccagata ctcatttatg attttgccaa ttttggtgtt
     3361 ctcaggttat cggagccagc accgctcttt gaccttgcca tgcttgcctt agatagtcca
     3421 gagagtggct ggacagagga agatggtccc aaagaaggac ttgctgaata cattgttgag
     3481 tttctgaaga agaaggctga gatgcttgca gactatttct ctttggaaat tgatgaggaa
     3541 gggaacctga ttggattacc ccttctgatt gacaactatg tgcccccttt ggagggactg
     3601 cctatcttca ttcttcgact agccactgag gtgaattggg acgaagaaaa ggaatgtttt
     3661 gaaagcctca gtaaagaatg cgctatgttc tattccatcc ggaagcagta catatctgag
     3721 gagtcgaccc tctcaggcca gcagagtgaa gtgcctggct ccattccaaa ctcctggaag
     3781 tggactgtgg aacacattgt ctataaagcc ttgcgctcac acattctgcc tcctaaacat
     3841 ttcacagaag atggaaatat cctgcagctt gctaacctgc ctgatctata caaagtcttt
     3901 gagaggtgtt aaggatcctg atatccagca cagtggcggc cgctcgagtc tagagggccc
     3961 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4021 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4081 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4141 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4201 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4261 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4321 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4381 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4441 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4501 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4561 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4621 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4681 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4741 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4801 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4861 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4921 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4981 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     5041 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     5101 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     5161 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5221 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5281 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5341 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5401 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5461 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5521 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5581 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5641 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5701 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5761 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5821 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5881 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5941 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6001 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6061 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6121 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6181 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6241 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6301 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6361 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6421 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6481 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6541 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6601 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6661 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6721 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6781 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6841 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6901 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6961 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7021 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7081 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7141 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     7201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8401 tctcccgatc ccctatggt
//
