LOCUS       V045690                 7656 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045690
VERSION     V045690
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..2336
                     /label="Pole2(NM_011133)"
                     /note="Pole2(NM_011133)"
                     /gene="Pole2"
     CDS             2343..2408
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2432..3020
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3086..3134
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3336..3764
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3778..4107
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4174..4968
                     /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    5144..5265
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5314..5330)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5338..5354
                     /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(5362..5392)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5407..5428
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5716..6304)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6475..7335)
                     /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(7336..7440)
                     /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 gccggagcgg ctgcggagcc
      781 gcacggtctc agctttcaag ctgcggggct tgctgctcag gagtgaagcc accaagtatc
      841 tgactgaagc tctccagtct gtcactgaat tggagctaga ggagaaccta ggaaagatca
      901 tcgatgcagt tgaaaagcag cccttgtcct cgaacatgat cgaacgatct gtcgtggagg
      961 cagctgtgca agagtgcagt cagtctatgg atgagactat agagcacgtt ttcaatatca
     1021 ttggagcttt tgatattcca cgttttatat ataattcaga aaggaaaaaa tttcttcctc
     1081 tatccatgac taatcatgct gcaccaacct tatttggaac agcaagagac aaagccgagc
     1141 tgtatcttga acgctatacc attttacacc agaggacaca cagacatgaa ttgttcactc
     1201 ctccggtgat aggctcacac cttgaagaaa gtgggagcaa gttccagctt aaaactatag
     1261 aaaccttact gggtagcaca agcaaagttg gtgacgtgat cgtcctgggg atgataaccc
     1321 agctgaaaga ggggaagttt ttcctggaag atcctactgg aacagtacaa ctggacctta
     1381 gtaaagctca gttccacagt ggcttataca ccgaggcctg cttcgtcctg gcagaaggtt
     1441 ggtttgaaga tcaagtgttt cacgccaatg cctttggatt tcctcccact gagccctcca
     1501 gcaccacaag ggcgtactat ggaaatatta atttttttgg agggccttct aatgcatcag
     1561 tgaagacgta tacaaagctg agacagctgg aagatgagaa caaggacgcc atgtttgtga
     1621 ttgtgtctga tgtttggttg gaccaagtgc aagtgttgga gaagtttcat gtaatgtttt
     1681 ctggttactc gccagcacct ccaacctgct ttattctgtg tggtaatttt tcatctgcac
     1741 catatggaaa aaatcaagtc caggctttga aagattccct aaaatctttg gcagacataa
     1801 tttgtgaata tccaagtatc caccaaagta gccgttttgt gtttgtgcct ggcccagagg
     1861 atcctggatt tggttccatt ttaccaaggc cgccactcgc tgaaagcatc acacaggaat
     1921 tcagacaaag ggttccgttt tcagttttca cgactaatcc ttgcagaatt cagtattgta
     1981 cacaagaaat aattatcttc cgtgaagact tagtgaataa aatgtgtaga aactgtgtcc
     2041 gttttcctag cagcagtttg gatattccta ctcacttcgt aaagaccatc ttatctcagg
     2101 gacatctggc tcctctccct ctttacgtat gccccgtcca ctgggcatat gattacacgt
     2161 tgagagtgta cccggtgccc gacctgctgg tcattgcaga caagtacgac ccgttcaccg
     2221 tgaccaacac cgattgcctc tgcataaacc ctggctcctt tccaagaagt ggatttgcat
     2281 tcaaagtttt ttacccttcc agtaaaacag tagaagacag caaacttcag ggctttggat
     2341 ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     2401 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     2461 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2521 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2581 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2641 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2701 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2761 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2821 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2881 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2941 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3001 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3061 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3121 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3181 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3241 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3301 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3361 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3421 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3481 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3541 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3601 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3661 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3721 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3781 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3841 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3961 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4021 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4081 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4141 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4201 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4261 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4321 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4381 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4441 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4501 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4561 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4621 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4681 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4741 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4801 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4861 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4921 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4981 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5041 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5101 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5161 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5221 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5281 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5881 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     5941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6121 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7441 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7501 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7561 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7621 ggcttgaccg acaattgcat gaagaatctg cttagg
//
