LOCUS       V035940                 9738 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035940
VERSION     V035940
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..4418
                     /label="REXO1(NM_020695)"
                     /note="REXO1(NM_020695)"
                     /gene="REXO1"
     CDS             4425..4490
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4514..5102
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5168..5216
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5418..5846
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5860..6189
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6256..7050
                     /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    7226..7347
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7396..7412)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7420..7436
                     /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(7444..7474)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7489..7510
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7798..8386)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8557..9417)
                     /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(9418..9522)
                     /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 ccaccatgct acgctccact ggcttcttcc
      781 gggccattga ctgcccctat tggtctgggg cgcccggggg gccctgccgg cggccctact
      841 gccacttccg gcaccgcggg gcccggggct ccggcgcgcc cggtgacggc ggagaggcgc
      901 cccccgcagc agggctgggt tacgacccct acaaccctga gctgcccaag ccccccgcgc
      961 agagggagaa tggcaccctg ggcctggggg aggagccgcg cccggatgtg ctggagttgg
     1021 agctggtcaa ccaggccatc gaggccgtgc gcagtgaggt ggagctggag cagcggcgct
     1081 accgggagct gctggagacg acccgtgagc accgctccgc cgaggccccc gccctggcgc
     1141 cccgcggccc caacgccagc cccactgtgg gcccggacga ggatgccttc ccactggcct
     1201 tcgactacag ccccggcagc cacggcctat taagccctga tgccggctac cagcccaccc
     1261 cactggccgc ccctgccgag ccgggcagca agtactcgct ggcgtccctg gacaggggtc
     1321 agggcagagg tggagggggt ggcggtgccc tggaatacgt ccccaaggct gtgagccagc
     1381 cccggcggca cagccgcccc gttcccagtg gcaagtacgt ggtggacaac tccaggccac
     1441 ccacagacct ggagtatgac cctctctcca actactcggc ccggcacctc agcagggcca
     1501 gctcccggga tgagcgggcc gccaagcggc cccggggctc ccgcggcagt gagccctaca
     1561 cacctgctcc caagaagctc tgtgacccct ttggcagttg cgatgcaagg ttctcagact
     1621 cagaagatga ggccgccacg gtcccaggta acgagcccac cacggccagc acccccaaag
     1681 ccagggccga ccctgagatc aaggccaccg ggcagccacc ctccaaagag ggcctggagg
     1741 ccgagggggg cggcctgcgg gagaccaagg agacggccgt gcagtgcgac gtgggggacc
     1801 tccagccgcc cccagccaag cccgcctccc cagcccaggt ccagtcctca caggatgggg
     1861 gctgccccaa ggagggaaaa cccaagaaga aaaaaaccgg ggccccacct gcccccagct
     1921 gcaaagacgg ggcccagggg aaggacaaga ccaaggacaa gggccgaggg cggcctgtgg
     1981 agaagccccg tgcggacaag aagggcccgc aggccagcag cccccggcgc aaggcagagc
     2041 ggccggaagg gaccaagaag aagccatctt cggccactcc tgtggccacc tcagggaaag
     2101 ggaggcctga ccggccagcg cggcggccga gccccacaag cggggactcc cgaccggcgg
     2161 ccggcagagg cccaccccgc cccctccagc tgcccgacag gaagagcacc aaggccccgt
     2221 cggggaagct agtggagcgg aaagcccgct cactagacga gggcgcctcc caggacgccc
     2281 ccaagctgaa gaagcgggcc ctgagccacg ccgacctctt tggggacgag agtgaggacg
     2341 aggccgcagg gccaggggtg ccgagcgtgt ggccctctgc cctccccagc ctcagctcgg
     2401 actcagactc cgactcagac tccagcctgg gcttcccgga ggcgcagggg ccgcccaagc
     2461 ggctcaaggc ctccccgccc ccctcccccg ccccatcctc ctcctcctcc tcctcctcct
     2521 ccacctccag cgcgggggcg gatgtggact actcggccct ggagaaggag gtggactttg
     2581 actccgaccc catggaggag tgcctgcgga tcttcaacga gtccaccagc gtcaagacgg
     2641 aggacagagg ccggctggcc cggcagcccc ccaaggaaga gaagagtgag gagaaggggc
     2701 tttcgggtct gaccactctg ttccccgggc agaagaggag gatctcccac ctttccaagc
     2761 aaggccagga ggtggagccc ccgaggaggg gtcccgcggt gcccccggcc cggcccccga
     2821 cggcgcagga ggtgtgctac ctgcgggccc agcaggcgca gagggcatcg gcgagcttgc
     2881 tgcaggcccc cgccaggctg gcagagaagt cgccctccgt ccacatttcc gcccctggcg
     2941 agaagaggag gatcgcccac atccccaacc cccgcctggc tgcagccccc acaggtgcca
     3001 agaggaccct tgcggccagc ggcagccagt cctccaacgg ccctgagcca ggtggccagc
     3061 agctgaaaac acgcacattg tcggggatgg cgtccaagac taccaccacc atcatcccta
     3121 agcgaatcgc ccacagtcca tccttacaga gtttaaagaa acccattatc cccaaagagt
     3181 ttgggggcaa agtccccacc gtcatccgcc agcgctatct caacctgttc atcgaggagt
     3241 gtctcaagtt ctgtacctcc aaccaggagg ccatagagaa ggcactgaac gaggagaagg
     3301 tggcctatga ccgcagcccc agcaagaaca tctacctgaa tgtggccgtg aacaccctca
     3361 agaagctcag gggcctggcc cccagcgctg tgcccggcct cagcaaaacc agtggccgca
     3421 gggttgtgtc ccacgaggtg gtgttggggg gcaggttggc cgccaagacc agcttctcgc
     3481 tcagccgtcc aagcagcccc cgggtggagg acctgaaagg ggctgccctg tacagccgcc
     3541 tcagggagta cctgctcacc caggaccagc tcaaggagaa cggctacccc ttcccgcacc
     3601 cagagcggcc cgggggcgca atcatcttca cagctgagga gaagaggccc aaggactctt
     3661 cctgcaggac ctgctgccgc tgtggcaccg agtacctcgt gtcctcttca ggccgctgca
     3721 tccgggacga ggagtgttat taccactggg gacggctgcg ccggaaccgg gtggccggag
     3781 gctgggagac ccagtacatg tgctgctcgg ctgccgccgg ctctgtcggc tgccaagtcg
     3841 caaagcaaca cgtgcaggat ggccggaagg agcgccttga gggcttcgtg aagacctttg
     3901 agaaagagct ctcaggagac acccacccgg ggatctacgc cctggactgc gagatgtcct
     3961 acaccacata tggcctggag ctgacgcgcg tcacggtggt cgacacggac gtgcacgtgg
     4021 tttatgacac cttcgtgaag cctgacaacg agatcgtgga ctacaacacc aggttttcgg
     4081 gggtgacgga ggctgacctt gccgacacaa gtgtcacgct gcgtgacgtc caggccgttc
     4141 tgctgagcat gttcagcgct gacaccatcc tcatcggaca cagcctggag agcgacctcc
     4201 tggccctgaa ggtcatccac agcaccgtgg tggacacgtc tgtgctcttc ccccaccgcc
     4261 tgggcctccc ctacaagcgg tccctgcgga acctcatggc cgactacctc agacagatca
     4321 tccaggacaa tgtggatggg cacagctcca gcgaggacgc cggcgcctgc atgcacctgg
     4381 tgatctggaa ggttcgagaa gacgccaaga ccaagcgagg atccgactac aaagaccatg
     4441 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     4501 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4561 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4621 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4681 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4741 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4801 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4861 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4921 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4981 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5041 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5101 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5161 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     5221 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5281 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5341 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     5401 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5461 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5521 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     5581 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5641 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5701 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5761 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     5821 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5881 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5941 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6001 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6061 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6121 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6181 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     6241 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6421 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6781 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     7021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     7081 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7141 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7201 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7261 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7321 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7381 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7441 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7501 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7981 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8221 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8521 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8581 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8641 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8701 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8761 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8821 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8881 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8941 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9001 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9061 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9121 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9181 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9241 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9301 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9361 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9421 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9541 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     9601 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     9661 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     9721 atgaagaatc tgcttagg
//
