LOCUS       V034782                 8011 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034782
VERSION     V034782
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             964..3501
                     /label="EXO1(NM_006027)"
                     /note="EXO1(NM_006027)"
                     /gene="EXO1"
     CDS             3508..3573
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3611..3835
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3881..4309
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4323..4652
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4719..5513
                     /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    5687..5808
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5857..5873)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5881..5897
                     /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(5905..5935)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5950..5971
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6259..6844)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7015..7875)
                     /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(7876..7980)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgggga tacagggatt gctacaattt atcaaagaag cttcagaacc catccatgtg
     1021 aggaagtata aagggcaggt agtagctgtg gatacatatt gctggcttca caaaggagct
     1081 attgcttgtg ctgaaaaact agccaaaggt gaacctactg ataggtatgt aggattttgt
     1141 atgaaatttg taaatatgtt actatctcat gggatcaagc ctattctcgt atttgatgga
     1201 tgtactttac cttctaaaaa ggaagtagag agatctagaa gagaaagacg acaagccaat
     1261 cttcttaagg gaaagcaact tcttcgtgag gggaaagtct cggaagctcg agagtgtttc
     1321 acccggtcta tcaatatcac acatgccatg gcccacaaag taattaaagc tgcccggtct
     1381 cagggggtag attgcctcgt ggctccctat gaagctgatg cgcagttggc ctatcttaac
     1441 aaagcgggaa ttgtgcaagc cataattaca gaggactcgg atctcctagc ttttggctgt
     1501 aaaaaggtaa ttttaaagat ggaccagttt ggaaatggac ttgaaattga tcaagctcgg
     1561 ctaggaatgt gcagacagct tggggatgta ttcacggaag agaagtttcg ttacatgtgt
     1621 attctttcag gttgtgacta cctgtcatca ctgcgtggga ttggattagc aaaggcatgc
     1681 aaagtcctaa gactagccaa taatccagat atagtaaagg ttatcaagaa aattggacat
     1741 tatctcaaga tgaatatcac ggtaccagag gattacatca acgggtttat tcgggccaac
     1801 aataccttcc tctatcagct agtttttgat cccatcaaaa ggaaacttat tcctctgaac
     1861 gcctatgaag atgatgttga tcctgaaaca ctaagctacg ctgggcaata tgttgatgat
     1921 tccatagctc ttcaaatagc acttggaaat aaagatataa atacttttga acagatcgat
     1981 gactacaatc cagacactgc tatgcctgcc cattcaagaa gtcatagttg ggatgacaaa
     2041 acatgtcaaa agtcagctaa tgttagcagc atttggcata ggaattactc tcccagacca
     2101 gagtcgggta ctgtttcaga tgccccacaa ttgaaggaaa atccaagtac tgtgggagtg
     2161 gaacgagtga ttagtactaa agggttaaat ctcccaagga aatcatccat tgtgaaaaga
     2221 ccaagaagtg cagagctgtc agaagatgac ctgttgagtc agtattctct ttcatttacg
     2281 aagaagacca agaaaaatag ctctgaaggc aataaatcat tgagcttttc tgaagtgttt
     2341 gtgcctgacc tggtaaatgg acctactaac aaaaagagtg taagcactcc acctaggacg
     2401 agaaataaat ttgcaacatt tttacaaagg aaaaatgaag aaagtggtgc agttgtggtt
     2461 ccagggacca gaagcaggtt tttttgcagt tcagattcta ctgactgtgt atcaaacaaa
     2521 gtgagcatcc agcctctgga tgaaactgct gtcacagata aagagaacaa tctgcatgaa
     2581 tcagagtatg gagaccaaga aggcaagaga ctggttgaca cagatgtagc acgtaattca
     2641 agtgatgaca ttccgaataa tcatattcca ggtgatcata ttccagacaa ggcaacagtg
     2701 tttacagatg aagagtccta ctcttttgag agcagcaaat ttacaaggac catttcacca
     2761 cccactttgg gaacactaag aagttgtttt agttggtctg gaggtcttgg agatttttca
     2821 agaacgccga gcccctctcc aagcacagca ttgcagcagt tccgaagaaa gagcgattcc
     2881 cccacctctt tgcctgagaa taatatgtct gatgtgtcgc agttaaagag cgaggagtcc
     2941 agtgacgatg agtctcatcc cttacgagaa gaggcatgtt cttcacagtc ccaggaaagt
     3001 ggagaattct cactgcagag ttcaaatgca tcaaagcttt ctcagtgctc tagtaaggac
     3061 tctgattcag aggaatctga ttgcaatatt aagttacttg acagtcaaag tgaccagacc
     3121 tccaagctac gtttatctca tttctcaaaa aaagacacac ctctaaggaa caaggttcct
     3181 gggctatata agtccagttc tgcagactct ctttctacaa ccaagatcaa acctctagga
     3241 cctgccagag ccagtgggct gagcaagaag ccggcaagca tccagaagag aaagcatcat
     3301 aatgccgaga acaagccggg gttacagatc aaactcaatg agctctggaa aaactttgga
     3361 tttaaaaaag attctgaaaa gcttcctcct tgtaagaaac ccctgtcccc agtcagagat
     3421 aacatccaac taactccaga agcggaagag gatatattta acaaacctga atgtggccgt
     3481 gttcaaagag caatattcca gctcgaggac tacaaagacc atgacggtga ttataaagat
     3541 catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgttta aacccgctga
     3601 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3661 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3721 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3781 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3841 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4081 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4201 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4261 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4321 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4381 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4441 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4501 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4561 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4621 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4681 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4741 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4801 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4861 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4921 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4981 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5041 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5101 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5161 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5221 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5281 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5341 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5401 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5461 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5521 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5821 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5881 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5941 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6001 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6061 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6781 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7981 cacatttccc cgaaaagtgc cacctgacgt c
//
