LOCUS       V018362                 9419 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018362
VERSION     V018362
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"
     CDS             671..4099
                     /label="MCM9(NM_017696)"
                     /note="MCM9(NM_017696)"
                     /gene="MCM9"
     CDS             4106..4171
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4195..4783
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4849..4897
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5099..5527
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5541..5870
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5937..6731
                     /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    6907..7028
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7077..7093)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7101..7117
                     /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(7125..7155)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7170..7191
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7479..8067)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8238..9098)
                     /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(9099..9203)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgccacc atgaatagcg atcaagttac actggttggt caagtgtttg agtcatatgt
      721 ttcggaatac cataagaatg atattcttct aatcttgaag gaaagggatg aagatgctca
      781 ttacccagtt gtggttaatg ccatgactct gtttgagacc aacatggaaa tcggggaata
      841 tttcaacatg ttccccagtg aagtgcttac aatttttgat agtgcactgc gaaggtcagc
      901 cttgacaatt ctccagtccc tttctcagcc tgaggctgtt tccatgaaac agaatcttca
      961 tgccaggata tcaggtttgc ctgtctgtcc tgagctggtg agggaacaca tacctaaaac
     1021 caaggatgtg ggacactttt tatctgtcac tgggacagtg attcgaacaa gtctggtgaa
     1081 ggttctggag tttgagcggg attacatgtg taacaaatgc aagcatgtgt ttgtgatcaa
     1141 ggctgacttt gagcagtatt acaccttttg ccggccatcc tcgtgtccca gcttggagag
     1201 ctgtgattcc tctaaattca cttgcctctc aggcttgtct tcgtctccaa ccaggtgtag
     1261 agattaccag gaaatcaaaa ttcaggaaca ggttcaaagg ctatctgttg gaagtattcc
     1321 acgatctatg aaggttattc tggaagatga cttagtggat agttgcaaat ctggtgatga
     1381 cctcactatt tacgggattg taatgcaacg gtggaagccc tttcagcaag atgtgcgctg
     1441 tgaagtggag atagtcctga aagcaaatta catccaagta aataatgagc agtcctcagg
     1501 gatcatcatg gatgaggagg tccaaaagga attcgaagat ttttgggaat actataagag
     1561 cgatcccttt gcaggaagga atgtaatatt ggctagcttg tgccctcaag tgtttggaat
     1621 gtatctagta aagcttgctg tggccatggt gctggctggt gggattcaaa ggactgatgc
     1681 tacaggaaca cgggtcagag gagaatctca tcttttattg gttggggatc ctggcacagg
     1741 gaaatctcag ttcctcaaat atgcagcaaa gattacacca agatctgtgc tgaccacagg
     1801 aattggatct actagtgcag gtctgacggt aactgctgta aaagactcag gagaatggaa
     1861 tttggaggct ggggcattag ttcttgcaga tgcgggcctt tgctgtattg atgagttcaa
     1921 tagcctcaaa gagcatgata ggaccagtat ccatgaagca atggagcaac aaaccataag
     1981 tgttgctaag gctggcctcg tgtgcaagct gaacacaagg accaccatcc tggcagcaac
     2041 gaaccccaaa ggccagtacg acccccagga gtccgtgtct gtgaacattg ccctcggcag
     2101 cccactctta agtcgatttg acctgatcct ggttttgctt gataccaaga atgaagactg
     2161 ggatcgtatc atttcctcct ttatcttaga aaataaaggt tacccaagca aatcagagaa
     2221 gctctggagc atggaaaaga tgaaaaccta tttctgcctc ataaggaatc tgcagcccac
     2281 actgtctgat gtgggcaatc aggttcttct ccggtactac cagatgcaaa ggcagagtga
     2341 ttgccggaac gctgcccgga ccaccattcg gctgttggaa agcttgatac gattagcaga
     2401 agctcatgct cgcctgatgt ttcgtgatac tgtaactctg gaagacgcta ttacggtggt
     2461 gtcagtcatg gagtcctcaa tgcagggagg tgcactgcta ggaggtgtga atgccctcca
     2521 cacttccttt cctgaaaacc ctggagagca gtaccagaga cagtgtgaac ttattctgga
     2581 aaagctagag ctgcagagcc tcttgagtga agagcttaga agacttgaaa ggttacagaa
     2641 tcagagtgtg caccaatccc aaccacgggt attggaggta gagactactc caggatcctt
     2701 gagaaatggt ccaggggaag aatcaaactt cagaacttca tcacagcagg aaatcaacta
     2761 tagcacacat atcttctctc ctggaggcag ccccgaggga agcccagttc tagatccccc
     2821 accgcatctg gagcctaata gatcaacaag taggaaacat tcagctcagc acaaaaataa
     2881 cagagatgac agtttagatt ggtttgattt catggcaact catcagagtg aacctaaaaa
     2941 cactgttgtt gtgtctcctc atcccaaaac atctggagaa aatatggctt cgaagatctc
     3001 taacagcaca tctcagggta aggagaagag tgagccaggc caaaggagca aagtggacat
     3061 tgggttgctt ccatcaccag gagagacagg tgttccatgg agggcagaca atgtggaaag
     3121 taacaagaaa aaaaggctag cactagattc tgaagcagca gtctctgctg ataaaccaga
     3181 ctcagtactg actcatcatg tccccaggaa cctgcagaag ctgtgcaaag agagggccca
     3241 gaagttgtgc agaaatagca ccagggtgcc tgcacagtgc acagtccctt cccatcctca
     3301 gtccactcct gtacatagcc cagacagaat gctggactca cccaaaagaa agagaccgaa
     3361 atcccttgcg caagtggaag agcctgcaat tgaaaatgtt aagcctccag gttcccctgt
     3421 ggccaaactg gcaaaattta ctttcaagca gaagtcaaaa ctgatccact cctttgaaga
     3481 tcacagccat gtgtcacctg gtgcaactaa aatagcagtt catagtccta aaatttccca
     3541 gcgtagaaca agaagagacg cagccttgcc ggtgaagcgt ccaggaaagt taacatctac
     3601 cccaggaaac cagatctcca gtcagccaca gggtgagaca aaggaggtgt cgcagcagcc
     3661 accagagaaa cacggaccaa gagagaaggt gatgtgtgcc cctgagaaga ggattattca
     3721 gcctgaatta gagcttggga acgagactgg gtgtgctcat cttacttgtg agggagacaa
     3781 aaaggaagag gtttcaggca gtaataaaag cggcaaggtt catgcctgca cattagccag
     3841 attggcaaac ttctgcttta ctcccccatc ggaatccaaa tcaaaatccc ctcctcctga
     3901 aaggaagaac cgaggtgaga gaggcccaag ctcccctcct acaaccacag ctccaatgcg
     3961 tgtcagtaaa aggaaatctt ttcagctccg tgggtccacc gagaaactga ttgtttccaa
     4021 agaatccctc ttcactttac cagaactagg tgatgaagca tttgattgtg actgggatga
     4081 agagatgaga aaaaagtcac tcgaggacta caaagaccat gacggtgatt ataaagatca
     4141 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     4201 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4261 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4321 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4381 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4441 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4501 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4561 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     4621 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4681 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4741 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     4801 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4861 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4921 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4981 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5041 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     5101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5221 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     5281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5461 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     5581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5641 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5821 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5881 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5941 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     6001 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     6061 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     6121 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6181 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6241 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6301 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6361 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6421 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     6481 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6541 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6601 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6661 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6721 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     6781 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6841 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6901 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7021 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7081 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7141 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7201 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     7681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     8881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9241 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     9301 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     9361 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
