LOCUS       V031415                 8252 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031415
VERSION     V031415
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3742
                     /label="MSH2(NM_000251)"
                     /note="MSH2(NM_000251)"
                     /gene="MSH2"
     CDS             3749..3814
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3852..4076
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4122..4550
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4564..4893
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4960..5754
                     /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    5928..6049
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6098..6114)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6122..6138
                     /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(6146..6176)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6191..6212
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6500..7085)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7256..8116)
                     /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(8117..8221)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggcggtgc agccgaagga
      961 gacgctgcag ttggagagcg cggccgaggt cggcttcgtg cgcttctttc agggcatgcc
     1021 ggagaagccg accaccacag tgcgcctttt cgaccggggc gacttctata cggcgcacgg
     1081 cgaggacgcg ctgctggccg cccgggaggt gttcaagacc cagggggtga tcaagtacat
     1141 ggggccggca ggagcaaaga atctgcagag tgttgtgctt agtaaaatga attttgaatc
     1201 ttttgtaaaa gatcttcttc tggttcgtca gtatagagtt gaagtttata agaatagagc
     1261 tggaaataag gcatccaagg agaatgattg gtatttggca tataaggctt ctcctggcaa
     1321 tctctctcag tttgaagaca ttctctttgg taacaatgat atgtcagctt ccattggtgt
     1381 tgtgggtgtt aaaatgtccg cagttgatgg ccagagacag gttggagttg ggtatgtgga
     1441 ttccatacag aggaaactag gactgtgtga attccctgat aatgatcagt tctccaatct
     1501 tgaggctctc ctcatccaga ttggaccaaa ggaatgtgtt ttacccggag gagagactgc
     1561 tggagacatg gggaaactga gacagataat tcaaagagga ggaattctga tcacagaaag
     1621 aaaaaaagct gacttttcca caaaagacat ttatcaggac ctcaaccggt tgttgaaagg
     1681 caaaaaggga gagcagatga atagtgctgt attgccagaa atggagaatc aggttgcagt
     1741 ttcatcactg tctgcggtaa tcaagttttt agaactctta tcagatgatt ccaactttgg
     1801 acagtttgaa ctgactactt ttgacttcag ccagtatatg aaattggata ttgcagcagt
     1861 cagagccctt aacctttttc agggttctgt tgaagatacc actggctctc agtctctggc
     1921 tgccttgctg aataagtgta aaacccctca aggacaaaga cttgttaacc agtggattaa
     1981 gcagcctctc atggataaga acagaataga ggagagattg aatttagtgg aagcttttgt
     2041 agaagatgca gaattgaggc agactttaca agaagattta cttcgtcgat tcccagatct
     2101 taaccgactt gccaagaagt ttcaaagaca agcagcaaac ttacaagatt gttaccgact
     2161 ctatcagggt ataaatcaac tacctaatgt tatacaggct ctggaaaaac atgaaggaaa
     2221 acaccagaaa ttattgttgg cagtttttgt gactcctctt actgatcttc gttctgactt
     2281 ctccaagttt caggaaatga tagaaacaac tttagatatg gatcaggtgg aaaaccatga
     2341 attccttgta aaaccttcat ttgatcctaa tctcagtgaa ttaagagaaa taatgaatga
     2401 cttggaaaag aagatgcagt caacattaat aagtgcagcc agagatcttg gcttggaccc
     2461 tggcaaacag attaaactgg attccagtgc acagtttgga tattactttc gtgtaacctg
     2521 taaggaagaa aaagtccttc gtaacaataa aaactttagt actgtagata tccagaagaa
     2581 tggtgttaaa tttaccaaca gcaaattgac ttctttaaat gaagagtata ccaaaaataa
     2641 aacagaatat gaagaagccc aggatgccat tgttaaagaa attgtcaata tttcttcagg
     2701 ctatgtagaa ccaatgcaga cactcaatga tgtgttagct cagctagatg ctgttgtcag
     2761 ctttgctcac gtgtcaaatg gagcacctgt tccatatgta cgaccagcca ttttggagaa
     2821 aggacaagga agaattatat taaaagcatc caggcatgct tgtgttgaag ttcaagatga
     2881 aattgcattt attcctaatg acgtatactt tgaaaaagat aaacagatgt tccacatcat
     2941 tactggcccc aatatgggag gtaaatcaac atatattcga caaactgggg tgatagtact
     3001 catggcccaa attgggtgtt ttgtgccatg tgagtcagca gaagtgtcca ttgtggactg
     3061 catcttagcc cgagtagggg ctggtgacag tcaattgaaa ggagtctcca cgttcatggc
     3121 tgaaatgttg gaaactgctt ctatcctcag gtctgcaacc aaagattcat taataatcat
     3181 agatgaattg ggaagaggaa cttctaccta cgatggattt gggttagcat gggctatatc
     3241 agaatacatt gcaacaaaga ttggtgcttt ttgcatgttt gcaacccatt ttcatgaact
     3301 tactgccttg gccaatcaga taccaactgt taataatcta catgtcacag cactcaccac
     3361 tgaagagacc ttaactatgc tttatcaggt gaagaaaggt gtctgtgatc aaagttttgg
     3421 gattcatgtt gcagagcttg ctaatttccc taagcatgta atagagtgtg ctaaacagaa
     3481 agccctggaa cttgaggagt ttcagtatat tggagaatcg caaggatatg atatcatgga
     3541 accagcagca aagaagtgct atctggaaag agagcaaggt gaaaaaatta ttcaggagtt
     3601 cctgtccaag gtgaaacaaa tgccctttac tgaaatgtca gaagaaaaca tcacaataaa
     3661 gttaaaacag ctaaaagctg aagtaatagc aaagaataat agctttgtaa atgaaatcat
     3721 ttcacgaata aaagttacta cgctcgagga ctacaaagac catgacggtg attataaaga
     3781 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg
     3841 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3901 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3961 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     4021 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4081 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     4141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4561 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4621 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4681 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4741 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4801 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4861 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4921 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4981 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     5041 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5101 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     5161 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5221 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5281 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5341 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5401 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5461 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     5521 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5581 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5641 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5701 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5761 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5821 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5881 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5941 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6001 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     6061 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6121 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     6181 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     6241 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6301 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6361 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6421 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6601 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6661 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6721 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6841 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6901 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6961 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7021 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8221 gcacatttcc ccgaaaagtg ccacctgacg tc
//
