LOCUS       V047103                10155 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047103
VERSION     V047103
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..4835
                     /label="Tnik(NM_026910)"
                     /note="Tnik(NM_026910)"
                     /gene="Tnik"
     CDS             4842..4907
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4931..5519
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5585..5633
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5835..6263
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6277..6606
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6673..7467
                     /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    7643..7764
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7813..7829)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7837..7853
                     /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(7861..7891)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7906..7927
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8215..8803)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8974..9834)
                     /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(9835..9939)
                     /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 gagcgactcc ccagctcgca
      781 gcctggatga aatcgatctc tccgccctga gggaccctgc agggatcttt gagttggtgg
      841 aacttgtcgg aaatggcacg tatggtcaag tttataaggg tcgtcatgtc aaaacgggcc
      901 agcttgctgc cattaaggtt atggatgtca caggggatga agaggaagaa atcaaacaag
      961 aaattaacat gttgaagaaa tattctcatc acaggaacat tgctacatac tacggtgctt
     1021 ttatcaaaaa gaaccctcct ggcatggatg accaactctg gttggttatg gagttctgtg
     1081 gtgctggctc tgtcactgac ctgatcaaga acacgaaagg caacacattg aaagaggagt
     1141 ggattgcata catctgcagg gagatcttac ggggcctgag tcacctgcac cagcacaaag
     1201 tgattcatcg agatatcaaa gggcagaacg tcttgttgac tgaaaatgca gaggttaagc
     1261 tagtggattt tggagtgagt gcccagcttg accgaactgt gggcaggagg aacacgttca
     1321 tcgggactcc ctactggatg gcaccagaag tcattgcctg tgatgagaac ccggatgcca
     1381 catatgattt caagagtgac ttgtggtctt tgggaatcac cgccattgag atggcagaag
     1441 gtgccccccc cctctgtgac atgcatccca tgagagccct cttcctcatc ccacggaacc
     1501 ctgcacctcg gctcaagtct aagaagtggt caaaaaaatt ccagtcattt atcgagagct
     1561 gcttggtaaa gaatcacagc cagcggccag ccacggagca gttgatgaag cacccattca
     1621 tacgagacca acctaatgag aggcaggtcc gcatccagct gaaggaccac attgatcgaa
     1681 caaagaagaa gcgaggagaa aaagatgaga ctgagtatga atacagcgga agtgaggaag
     1741 aagaggaaga gaatgactct ggggaaccca gctccattct gaacctacca ggggagtcaa
     1801 cactgcgaag ggacttcctg agactgcagc tggccaacaa ggagcgctca gaggccctgc
     1861 ggcgccaaca gctggagcag cagcagcggg agaatgaaga acacaagcgg cagctactgg
     1921 ctgagcgcca gaagcgcatc gaagagcaga aggagcaaag gcggaggctg gaggagcaac
     1981 aaaggcgaga aaaagagctt cggaaacagc aggagcggga acagcgccgg cactacgaag
     2041 aacagatgcg tcgggaggag gagaggaggc gtgccgaaca tgagcaggaa tacatcaggc
     2101 ggcagctaga agaagagcaa aggcagttag agatcttaca gcagcagcta ctgcatgagc
     2161 aagctctact tctggaatat aagcgcaaac aattggaaga acagagacaa gcagaaagac
     2221 tgcagaggca gctaaagcaa gagcgggact atctggtttc cctccagcat cagcggcagg
     2281 agcagaggcc cctggagaag aagccactgt accattacaa ggagggcatg agtcctagtg
     2341 agaagccggc ctgggccaag gaggtagaag aacgctcaag actcaaccga cagagttcac
     2401 ctgccatgcc tcacaaggtt gccaacagga tctcggaccc caacctgccc ccaagatcag
     2461 agtccttcag cattagtggg gttcagcctg caaggacacc cccaatgctc agacctgttg
     2521 acccccagat cccgcagctg gtagctgtca aatcccaggg acctgccttg acggcctccc
     2581 agtcagtaca tgagcaaccc acaaagggcc tgtctgggtt ccaggaggct ctgaatgtga
     2641 cctctcaccg ggtcgagatg ccacgccaga actcggatcc cacctcagaa aaccctcctc
     2701 tccccacgag aattgagaag tttgacagaa gctcttggtt acgacaggaa gaagacattc
     2761 caccaaaggt gcctcaaaga acaacttcta tatccccagc actagccaga aagaattccc
     2821 ctggcaatgg cagtgctctg ggccccagac ttggatctca gcccatcaga gcaagcaacc
     2881 ctgatctgcg caggacagag ccagtcctgg agagttccct gcagcggaca agcagtggca
     2941 gttcctccag ctccagcact cccagctccc agcccagctc ccaaggaggc tctcaacctg
     3001 gctcccaagc aggatctagt gagcggtcca gagtgcgggc caacagtaag tccgaaggat
     3061 cacccgtgct cccccatgag ccttccaagg tgaaaccaga agaatccaga gacatcacac
     3121 ggcccagtcg gccagctagc tataagaaag ctatagatga ggatctgacg gcattagcca
     3181 aagaattacg agaactccgc attgaagaaa caaaccgccc cctgaagaaa gtgactgatt
     3241 actcttcctc cagcgaggag tcggagagca gtgaggaaga agaggaagat ggagagagtg
     3301 agacacatga cgggacggtg gctgtcagtg acatacccag actaataccc accggagctc
     3361 cagggaacaa tgagcagtac aacatgggga tggtcgggac acatgggctg gaaacttcgc
     3421 atgcggacac ctttggcggc agcatttcaa gagaaggaac cttgatgatc agagagacgg
     3481 ctgaagagaa gaagcgatct ggccacagtg acagtaatgg attcgccggt cacatcaatc
     3541 tcccagacct tgtacagcag agccattcgc cagctggaac tcccactgag gggctgggcc
     3601 gcgtctccac tcattcccag gagatggact ctggggctga atatggtata gggagcagca
     3661 ccaaagcctc tttcaccccc ttcgtggacc ctcgagtgta ccagacatcg cccactgatg
     3721 aagatgaaga ggatgatgag tcttcagctg ctgccctgtt tactagcgaa cttcttaggc
     3781 aagaacaggc caaactcaat gaagcgagga agatttcagt ggtaaatgtg aacccaacaa
     3841 acattcgccc tcatagtgac acaccggaaa tcagaaaata caagaaacgc ttcaattcag
     3901 aaatactttg tgcagctcta tggggtgtga accttctggt ggggactgaa aatggcctga
     3961 tgcttttgga cagaagtggc caaggcaaag tctacaacct aatcaaccgg aggcggtttc
     4021 agcagatgga tgtgctagaa ggactaaatg ttcttgtcac gatatcagga aagaagaaca
     4081 agctccgtgt gtactatctc tcatggttaa gaaacagaat cctgcacaat gacccagaag
     4141 tggaaaagaa gcagggctgg atcactgtcg gtgacttgga aggctgcatc cattacaaag
     4201 tcgttaaata tgaaagaatc aagttcctgg tgattgcctt aaagaatgca gtagagatat
     4261 atgcgtgggc ccctaaacct taccataagt tcatggcatt taagtctttt gcagatcttc
     4321 agcataagcc tctgctcgtt gacctcacag tagaagaagg tcaaaggtta aaggtcatat
     4381 ttggctcaca cactggtttc catgtaattg atgttgattc tggaaactcc tacgatatct
     4441 atataccatc ccatattcag ggcaatatca ctcctcatgc tatcgtcatc ttgcctaaaa
     4501 cagatggaat ggagatgctt gtctgctatg aggatgaagg ggtgtacgtg aacacctacg
     4561 gccggatcac taaggatgtg gtgctccaat ggggagaaat gcccacatct gtggcctaca
     4621 ttcattccaa tcagataatg ggctggggcg agaaagctat tgagatccgg tcagtggaaa
     4681 caggacattt ggatggagtg tttatgcata aacgagctca aaggttaaag tttctatgtg
     4741 aaagaaatga taaggtattt tttgcatccg tgcgatctgg aggaagtagc caagtgtttt
     4801 tcatgaccct caacagaaat tccatgatga actggggatc cgactacaaa gaccatgacg
     4861 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     4921 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4981 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5041 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5101 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5161 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5221 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5281 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5341 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5401 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5461 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5521 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5581 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     5641 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5701 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5761 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     5821 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     6001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6181 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     6241 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     6301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6541 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6601 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     6661 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     6721 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     6781 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     6841 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     6901 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     6961 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     7021 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     7081 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     7141 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     7201 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     7261 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     7321 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     7381 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     7441 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     7501 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7561 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7621 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7681 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7741 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7801 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7861 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7921 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7981 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     8041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     8101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     8161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     8221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8401 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8641 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8821 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8881 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8941 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     9001 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     9061 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     9121 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     9181 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     9241 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     9301 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     9361 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     9421 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9481 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9541 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9601 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9661 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9721 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9781 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9961 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
    10021 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
    10081 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
    10141 aagaatctgc ttagg
//
