LOCUS       V044655                 9024 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044655
VERSION     V044655
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..3704
                     /label="Zmym1(NM_026670)"
                     /note="Zmym1(NM_026670)"
                     /gene="Zmym1"
     CDS             3711..3776
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3800..4388
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4454..4502
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4704..5132
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5146..5475
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5542..6336
                     /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    6512..6633
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6682..6698)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6706..6722
                     /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(6730..6760)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6775..6796
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7084..7672)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7843..8703)
                     /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(8704..8808)
                     /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 tcctgcagtt ccaggcacag
      781 ctgttcgact ctcctgtgcc aaatgtaaaa gagtttttca ggagggacag acagcttttc
      841 agaggaaagg gtctgctgag cttttctgct caagactatg cattgatggg tacccgtcac
      901 ctgctgtgtc tccagctcct ctcaaaagga tgtgcttaaa ttgctcaaga gacattttaa
      961 gtctaaagga tatgaagagt gtccagttgg aggatagtag ttgtgacaaa aacttttgta
     1021 gccaatcttg tcttacatca tatgaagaaa aaagaaaacc aggtgttacc atatatactg
     1081 atagcagttt gcccaagtgc agcatgtgtc agaagacatc tactaaaccg cccagaatac
     1141 ttagatcttc cccgtgcaaa tcactgcagc tgtcagatga gatgattgcg attactaacg
     1201 acttggggaa cgtagaggtt tactgctctg cttgttcttc ttcatacaac agtgctgtga
     1261 tggagtctcc ttcagttaat gtatccatgg tgcacagttc ttcaaaagag aatctctgtc
     1321 caaagaaata tccagttata agcaatataa tgtcgttagt aaataatcat gttgacctgg
     1381 tcgtgaacac tgatctctta caaggtacac tttcttctct accgacagat gtcatcatga
     1441 gcaactcagc cagtgagtcc agcagtggtg ttggtaatag tgcggaccag ccaggcctcc
     1501 tcccaccttg ctcggtgctc tgtgaggaca cagctgctcc cggtgaggga gtagagaaca
     1561 acagtgagaa atccatggaa ggaaatgata ggctgtgtct ctcaaagtcc acattaacag
     1621 aacagaacat taagacagag cctccgggag agccctggag cccagctctc cagcatctgc
     1681 atcccagccc tgcagacaat atcgcattct gctattcttg ctacttgttc ttccagaaaa
     1741 aatttagctg tcggagagaa gcttttgcaa cccaaggaac tgctgactgg gagaaaatgc
     1801 tgggaaattt cagaaagcat gaagaaagtg acgtgcactt aaagtcactg caattttgga
     1861 ggcaatacca gtttctcgat gctattccgt gtgacaattc atctgtttat tcaaaacaga
     1921 ttgaaggaaa taaaaagtac ctaaggctca tcattgagaa cattttattt cttggaaaac
     1981 agtgcttgct gttaaaagga aatgaccagt ctgtttcacc catgaataaa ggcagttttt
     2041 tggaattgct agaacttaga gcaagagata gaggagaggt atttcagctc atgagttccc
     2101 acgtggactt ctatagcagc acgcttgtcc aggaggagat tattgaaatg ataaaggatg
     2161 aaatgttgca agacatcgtg agtgagatca acgcctcctc agccttttca ataatctgcg
     2221 aggagatagc tggtagtgcc gctgagagac agctttccgt gtgtgtgagg tacccacaaa
     2281 agacgccaag tgctgtcttg gttaaggaaa ggttcctggg gtttgtgact gccgaggagg
     2341 tgactggggt ccacgtgcat gggcggatca aagcttacct gcagcaggtt ggagttgatt
     2401 ttaataaaat atgtggccaa gcctatgata gtgccacaaa ttttacggta aaattgaatg
     2461 aagttgtagc agaattcaag aaggaagagc caagagctct ctatgtgcat tgtcacgccc
     2521 attttttgga gttagcggtg attagcttct gtaaggaggt gaaggagctc cgcgaggcac
     2581 tacacactct ctgctctttg ttcaacacag ttcatgtgtc tgaagaaatg ttggcaagtt
     2641 tgcaaaccat gtgtaagcta agtctaaata aaacttgtaa gaaacacacg tcacagtcgt
     2701 gttggccagc ccacgaccat ttgctgctgg ccgtgacgga ctgccttccg gaggtcgttg
     2761 aggtgctggc gtcagtgtca cgccggtgcg caggcacagc tgtggctgcc gagctggagg
     2821 acctgttggc catggttacc aagtttgaat ttatattttg tttgaaattt ctttatcgag
     2881 tactcagtat cacagggatt ctttctagag agtttcaaag tgaaactgtt gacatctttt
     2941 ctttgttttc aaaaatagaa gcaattttgg aatgtttatc ttctgaaagg aatgatacat
     3001 actttaaaaa tatctgggat ggagcagatg aaatatgtaa aaagataacc agtaagggtt
     3061 ttgatgttga gaaaccctct tttcaaaaaa gaagaaaagt tcaaagaaca gtagatcctg
     3121 gcaactcaga gcatatgttt tttcctactt cagcagatga acaatataaa tttaatattt
     3181 actaccaagg tctggacacg gtattaaaac atttaaagtt atatttttca gagtttgact
     3241 attgtaaaat gaaacaaatt tcagagctgt tacttaagtg gaatgagcca ttaaatgaag
     3301 caacagccaa gcacatccag gaattctata agcttgatgc agacatcatt cccgagctcc
     3361 gattttaccg gcagtatgca ggtttcaact tcatcgcaga ggatgggtcc ttaagcttca
     3421 ttgacctagg ctctttgttt attcagcatg ctctccacaa taatattcct tgtatcacga
     3481 agctgttgca tattgcttta tcttggccaa ttacttcagc aaataatgag aagtcatttt
     3541 ccacgctgtc tcatcttaaa acatacttac ttcgtaccat gggacaagag aaacttagta
     3601 gcctggctct gatagctgtt gagcaggaac tggtaaataa actgatggaa cctgaaagac
     3661 tggggggaat tgtcgaaaag ttcattagtc aaatgaaaga tacgggatcc gactacaaag
     3721 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     3781 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3841 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3901 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3961 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     4021 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     4081 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     4141 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     4201 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     4261 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4321 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4381 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     4441 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     4501 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4561 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     4621 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     4681 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4741 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4801 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4861 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4921 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4981 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5041 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     5101 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     5161 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5221 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5281 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5341 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5401 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5461 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5521 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5581 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5641 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5701 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5761 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5821 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5881 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5941 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6001 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6061 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6121 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6181 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6241 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6301 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     6361 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6421 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6481 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6541 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6601 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6661 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6721 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6781 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6841 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6901 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6961 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7021 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7081 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7141 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7201 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7261 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7321 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7381 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7441 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7501 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7561 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7621 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7681 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7741 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7801 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7861 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7921 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7981 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8041 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8101 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8161 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8221 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8281 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8341 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8401 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8461 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8521 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8581 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8641 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8701 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8761 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8821 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     8881 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     8941 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     9001 aattgcatga agaatctgct tagg
//
