LOCUS       V045631                 8271 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045631
VERSION     V045631
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2951
                     /label="Xrcc5(NM_009533)"
                     /note="Xrcc5(NM_009533)"
                     /gene="Xrcc5"
     CDS             2958..3023
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3047..3635
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3701..3749
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3951..4379
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4393..4722
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4789..5583
                     /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    5759..5880
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5929..5945)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5953..5969
                     /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(5977..6007)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6022..6043
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6331..6919)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7090..7950)
                     /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(7951..8055)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc gtggtccggt aataaggcag
      781 ctgttgtgct gtgtgtggat gtgggggttg ccatgggtaa ctcctttcct ggtgaagaat
      841 ctccaattga acaagcaaag aaagtgatga ctatgtttgt ccaacgacag gtattttcgg
      901 agagcaaaga tgagattgcg ttagtcctct atggcacaga tggcactgat aatgcccttg
      961 ctggcaagga ccagtatcag aacatcacag tgtgcagaca cctgatgcta ccagattttg
     1021 atttgctgga agacatcgga aacaaaatcc agccaagttc ccagcaagcc gacttcctgg
     1081 acgccctgat tgtgtgcatg gatttgattc agcgtgaaac catagggaag aagtttggga
     1141 agaagcatat tgaagtgttc actgacctca gcagcccgtt cagccaggac caactggacg
     1201 ttataatttg taacttgaag aagtctggca tctctctgca gttctttttg ccttttccaa
     1261 tcgacaagaa tggcgagcct ggcgagagag gggacctgga ctctggcttg gaccacctca
     1321 aaccctcctt ccctcaaaaa ggacttactg agcagcaaaa ggaaggcatc cgcatggtga
     1381 caagggtgat gctgtcctta gaaggcgaag acgggctgga tgaaatttat tccttcagtg
     1441 agagtctacg acaactgtgc gtctttaaga agattgagag gcgttccatg ccctggccct
     1501 gccaactgac cattggcccc aacttgtcta taaagattgt agcctataaa tcgattgtac
     1561 aggagaaatt taagaagagt tgggtagttg tggacgcaag aacgctaaag aaggaagata
     1621 tacaaaaaga gactgtttat tgcctaaatg acgatgatga aactgaagtt tccaaagagg
     1681 acactattca agggtaccgc tacggaagtg atatcattcc tttttctaaa gtggatgagg
     1741 aacaaatgaa atataaatct gaggggaagt gcttctctgt tttgggattt tgtaaatctt
     1801 ctcaggttca tagaagattc ttcatgggac atcaagttct aaaggtcttt gcagcaaagg
     1861 atgatgaggc ggcagctgtt gccctttcct ctctcgttca tgctttggat gagttaaaca
     1921 tggtcgccat cgtccgatat gcttatgaca aaagatctaa tcctcaagtt ggtgtagcct
     1981 tcccttacat taaggacgcc tatgagtgtt tagtttatgt gcagctgcct ttcatggaag
     2041 acttgcggca atacatgttt tcctcgctga aaaacaataa gaagtgcact cccacagagg
     2101 cccagctgag cgctattgat gatctgattg attccatgag cttggtaaag aaaaacgagg
     2161 aagaagacat tgttgaagat ttgtttccaa cctccaaaat tccaaatcct gaatttcagc
     2221 gattgtacca gtgtctgctg catagagcct tacatctcca ggagcggctg cccccgattc
     2281 agcagcacat tttgaatatg ctggatcccc ccactgagat gaaagcaaaa tgtgagagtc
     2341 cactctctaa agtaaagacc cttttccctc tcacagaagt catcaagaaa aagaaccaag
     2401 tgactgctca ggacgttttc caagacaatc atgaagaggg gcccgctgct aaaaaatata
     2461 aaactgagaa agaagaagat cacatcagca tctccagcct ggcagaaggg aacatcacca
     2521 aggttggaag tgtgaatcct gttgaaaact tccgtttcct agtaagacag aagattgcca
     2581 gctttgagga agcgagtctc cagttaataa gtcacatcga acaatttttg gataccaatg
     2641 aaacactgta ttttatgaaa agtatggact gcatcaaagc tttccgggag gaggccattc
     2701 agttttcaga agaacagcgc ttcaacagct tcctggaagc ccttcgagag aaagtggaaa
     2761 ttaagcaatt aaatcatttc tgggaaattg ttgttcagga tggagttact ctgatcacca
     2821 aagacgaagg cccaggaagc tctatcacag ctgaggaagc cacaaagttt ctggccccca
     2881 aagacaaagc aaaagaagat acaacgggac ctgaagaagc tggtgatgtg gatgatttac
     2941 tggacatgat aggatccgac tacaaagacc atgacggtga ttataaagat catgacatcg
     3001 actacaagga tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga
     3061 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3121 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3181 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3241 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3301 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3361 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3421 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3481 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3541 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3601 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3661 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     3721 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     3781 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     3841 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     3901 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     3961 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4021 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4081 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     4141 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4201 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4261 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4321 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4381 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4441 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4501 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4561 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4621 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4681 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4741 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4801 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4861 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4921 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4981 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5041 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5101 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5161 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5221 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5281 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5341 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5401 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5461 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5521 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5581 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5641 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5701 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5761 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5821 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5881 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5941 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6001 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6061 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7201 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8101 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     8161 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     8221 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
