LOCUS       V046651                 8565 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046651
VERSION     V046651
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..3245
                     /label="Ttk(NM_009445)"
                     /note="Ttk(NM_009445)"
                     /gene="Ttk"
     CDS             3252..3317
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3341..3929
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3995..4043
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4245..4673
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4687..5016
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5083..5877
                     /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    6053..6174
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6223..6239)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6247..6263
                     /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(6271..6301)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6316..6337
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6625..7213)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7384..8244)
                     /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(8245..8349)
                     /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 ccaccatgga ggctgaagag ttaattggca
      781 gcagtgtgac gattgattcc atcatgagca aaatgagaga tattaaaaat aagataaatg
      841 aagactgtac tgatgagcta agcttgtcta aaatctgtgc cgatcacacg gaaactgtta
      901 accaaattat gagggttggg aacaccccag agaactggtt gaatttcttg ctgaaactag
      961 agaaaaacag ctcacctcta aatgacgatc ttttaaataa gctgattggt cggtatagtc
     1021 aagcgattga agtacttcct ccagataaat acggccagaa tgagagcttt gctcgaatac
     1081 aagtgagact tgctgaacta aaagctattc aagagcctga tgatgcccgt gactacttcc
     1141 agatggccag ggaaaactgc aagaagtttg cttttgtgca cgtatctttt gcacagtttg
     1201 aactgtctca aggcaatctt aaaaaaagtg agcagcttct tcataaagct gtagagactg
     1261 gggcggtgcc gctgcagatg ctggagacgg ccatgcgtaa cttacacctc cagaaaaagc
     1321 agctgcttcc ggaggaggac aagaagagtg tgtcagcatc gacagtacta agtgcccaag
     1381 agccgttctc cagctcactt ggaaatgtac agaatcggag catcagctgt gagtccagag
     1441 gacaggctgg ggcagccagg gttttatatg gagagaactt gcctccacaa gatgccgaag
     1501 tgaggcatca aaaccccttc aagcagactc acgcagctaa acggtcatgc ccctttggaa
     1561 gagtcccagt caatcttcta aacagcccag atttctatgt gaagacagat agctcagctg
     1621 tgacacagtt aacaacaaga aaggggtcag gaccagaccg agacgcgatt ctgcccggct
     1681 ccagaccacg tggcagtgat tcctatgaac tgagaggttt aaagcccatt caaactatct
     1741 atttgaaaga ctctttggtg tccaatgaaa agagttctga acttatgtct gatttaatag
     1801 ccttgaagag taaaacagat tcaagtctaa caaaattgga agaaactaag ccagagattg
     1861 cagaaagaag gcccatgcag tggcagtcta ccagaaagcc cgagtgtgtg ttccagaacc
     1921 ctgctgcctt tgcacccctg cggcacgttc cagatgtcac cccgaaggct gacaaagagt
     1981 caccaccaat atcagttcct aaatggcttg atccaaagtc tgcttgtgag acacctagta
     2041 gcagctcctt ggatgattac atgaaatgtt ttaagactcc agttgtaaag aatgactttc
     2101 cacctgcctg tccatcatca acaccttaca gccagcttgc ccgcctccag cagcaacagc
     2161 agcagggact cagcactcct cttcaaagct tgcagatttc aggttcttca tcaataaatg
     2221 aatgcatttc agttaacgga agaatttatt ccatattaaa gcagataggc agtggaggtt
     2281 ccagtaaggt gtttcaggta ttgaatgaga aaaaacagat aaacgctatc aaatatgtga
     2341 acctagaaga cgccgatagc caaactattg agagctaccg caacgagata gcgtttttga
     2401 acaaactaca gcaacacagt gataagatca tccgcctcta tgattatgaa atcaccgagc
     2461 agtacatcta catggtaatg gaatgtggaa acattgacct aaatagttgg cttaaaaaga
     2521 aaaaatccat caatccatgg gaacgcaaga gctactggaa aaacatgttg gaggcagtac
     2581 acataatcca tcagcatggt attgttcata gtgatctgaa gcctgctaac tttgtgatag
     2641 tggatggaat gctaaagcta attgattttg ggattgcaaa ccaaatgcag ccagacacaa
     2701 caagcattgt taaagattct caggttggca cagttaacta tatggcccca gaagcaatca
     2761 gagacatgtc ttcttcaaga gaaaattcga aaatcaggac caaggtaagt cccagaagtg
     2821 atgtctggtc cttggggtgc attttgtact acatgactta tgggaggacg ccatttcagc
     2881 acatcatcaa tcaggtctct aaactgcacg ccataatcaa ccctgctcat gagattgaat
     2941 ttcccgagat ttcggaaaaa gatcttcgag acgtgttaaa gtgctgttta gtgaggaacc
     3001 ctaaagagag gatatctatc cctgagcttc tcacacatcc gtatgttcaa attcagcccc
     3061 atccaggcag ccaaatggct aggggagcca ctgatgaaat gaaatatgtg ttgggtcaac
     3121 ttgttggtct gaattctcct aactccatct tgaaaactgc aaaaactttg tatgaacgtt
     3181 ataattgtgg tgaaggtcaa gattcttcgt catccaagac ttttgacaaa aagagagaaa
     3241 gaaagggatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca
     3301 aggatgacga tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa
     3361 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3421 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3481 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3541 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3601 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3661 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3721 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     3781 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3841 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3901 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     3961 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4021 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4081 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4141 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4201 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4261 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4321 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4381 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4441 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4501 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4561 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4621 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4681 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4741 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4801 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4861 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4921 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4981 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5041 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5101 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5161 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5221 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5281 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5341 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5401 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5461 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5521 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5581 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5641 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5701 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5761 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5821 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5881 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5941 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6001 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6061 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6121 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6181 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     6361 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     6421 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     6841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8401 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     8461 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     8521 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
