LOCUS       V020530                 7718 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020530
VERSION     V020530
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..3208
                     /label="MLH1(NM_000249)"
                     /note="MLH1(NM_000249)"
                     /gene="MLH1"
     CDS             3215..3280
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3318..3542
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3588..4016
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4030..4359
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4426..5220
                     /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    5394..5515
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5564..5580)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5588..5604
                     /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(5612..5642)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5657..5678
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5966..6551)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6722..7582)
                     /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(7583..7687)
                     /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 atgtcgttcg tggcaggggt
      961 tattcggcgg ctggacgaga cagtggtgaa ccgcatcgcg gcgggggaag ttatccagcg
     1021 gccagctaat gctatcaaag agatgattga gaactgttta gatgcaaaat ccacaagtat
     1081 tcaagtgatt gttaaagagg gaggcctgaa gttgattcag atccaagaca atggcaccgg
     1141 gatcaggaaa gaagatctgg atattgtatg tgaaaggttc actactagta aactgcagtc
     1201 ctttgaggat ttagccagta tttctaccta tggctttcga ggtgaggctt tggccagcat
     1261 aagccatgtg gctcatgtta ctattacaac gaaaacagct gatggaaagt gtgcatacag
     1321 agcaagttac tcagatggaa aactgaaagc ccctcctaaa ccatgtgctg gcaatcaagg
     1381 gacccagatc acggtggagg acctttttta caacatagcc acgaggagaa aagctttaaa
     1441 aaatccaagt gaagaatatg ggaaaatttt ggaagttgtt ggcaggtatt cagtacacaa
     1501 tgcaggcatt agtttctcag ttaaaaaaca aggagagaca gtagctgatg ttaggacact
     1561 acccaatgcc tcaaccgtgg acaatattcg ctccatcttt ggaaatgctg ttagtcgaga
     1621 actgatagaa attggatgtg aggataaaac cctagccttc aaaatgaatg gttacatatc
     1681 caatgcaaac tactcagtga agaagtgcat cttcttactc ttcatcaacc atcgtctggt
     1741 agaatcaact tccttgagaa aagccataga aacagtgtat gcagcctatt tgcccaaaaa
     1801 cacacaccca ttcctgtacc tcagtttaga aatcagtccc cagaatgtgg atgttaatgt
     1861 gcaccccaca aagcatgaag ttcacttcct gcacgaggag agcatcctgg agcgggtgca
     1921 gcagcacatc gagagcaagc tcctgggctc caattcctcc aggatgtact tcacccagac
     1981 tttgctacca ggacttgctg gcccctctgg ggagatggtt aaatccacaa caagtctgac
     2041 ctcgtcttct acttctggaa gtagtgataa ggtctatgcc caccagatgg ttcgtacaga
     2101 ttcccgggaa cagaagcttg atgcatttct gcagcctctg agcaaacccc tgtccagtca
     2161 gccccaggcc attgtcacag aggataagac agatatttct agtggcaggg ctaggcagca
     2221 agatgaggag atgcttgaac tcccagcccc tgctgaagtg gctgccaaaa atcagagctt
     2281 ggagggggat acaacaaagg ggacttcaga aatgtcagag aagagaggac ctacttccag
     2341 caaccccaga aagagacatc gggaagattc tgatgtggaa atggtggaag atgattcccg
     2401 aaaggaaatg actgcagctt gtaccccccg gagaaggatc attaacctca ctagtgtttt
     2461 gagtctccag gaagaaatta atgagcaggg acatgaggtt ctccgggaga tgttgcataa
     2521 ccactccttc gtgggctgtg tgaatcctca gtgggccttg gcacagcatc aaaccaagtt
     2581 ataccttctc aacaccacca agcttagtga agaactgttc taccagatac tcatttatga
     2641 ttttgccaat tttggtgttc tcaggttatc ggagccagca ccgctctttg accttgccat
     2701 gcttgcctta gatagtccag agagtggctg gacagaggaa gatggtccca aagaaggact
     2761 tgctgaatac attgttgagt ttctgaagaa gaaggctgag atgcttgcag actatttctc
     2821 tttggaaatt gatgaggaag ggaacctgat tggattaccc cttctgattg acaactatgt
     2881 gccccctttg gagggactgc ctatcttcat tcttcgacta gccactgagg tgaattggga
     2941 cgaagaaaag gaatgttttg aaagcctcag taaagaatgc gctatgttct attccatccg
     3001 gaagcagtac atatctgagg agtcgaccct ctcaggccag cagagtgaag tgcctggctc
     3061 cattccaaac tcctggaagt ggactgtgga acacattgtc tataaagcct tgcgctcaca
     3121 cattctgcct cctaaacatt tcacagaaga tggaaatatc ctgcagcttg ctaacctgcc
     3181 tgatctatac aaagtctttg agaggtgtct cgaggactac aaagaccatg acggtgatta
     3241 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     3301 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3361 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3421 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3481 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3541 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3661 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3721 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3781 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3901 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3961 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4021 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4141 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4201 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4261 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4321 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4381 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4441 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4501 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4561 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4621 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4681 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4741 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4801 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4861 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4921 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4981 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5041 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5101 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5161 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5221 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5281 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5341 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5521 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5581 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5641 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5701 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5761 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5821 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5881 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5941 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6481 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6841 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7681 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
