LOCUS       V044956                 7395 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044956
VERSION     V044956
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..2075
                     /label="Vrk1(NM_011705)"
                     /note="Vrk1(NM_011705)"
                     /gene="Vrk1"
     CDS             2082..2147
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2171..2759
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2825..2873
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3075..3503
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3517..3846
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3913..4707
                     /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    4883..5004
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5053..5069)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5077..5093
                     /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(5101..5131)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5146..5167
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5455..6043)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6214..7074)
                     /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(7075..7179)
                     /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 ccaccatgcc ccgtgtaaaa gcagctcagg
      781 ctggaagacc cggacctgcg aagaggcgcc tcgcagagca gtttgccgct ggagaggtcc
      841 taaccgacat gtctaggaag gagtggaaac taggattgcc cattggccaa ggtggctttg
      901 gctgcatcta tctggcggac acaaattctt ccaaaccggt tggcagtgac gcgccctgtg
      961 ttgtgaaagt ggaacccagt gacaatggac ctcttttcac ggaattaaag ttctaccaga
     1021 gggctgctaa accagagcaa attcagaaat ggattcgtac acataaattg aagtaccttg
     1081 gtgttcctaa gtattgggga tctggtctac atgataaaaa tggaaaaagt tacaggttta
     1141 tgataatgga ccgctttggg agtgaccttc agaaaatata tgaagcaaat gccaaaaggt
     1201 tttctcggaa aactgtattg cagctaagct taagaattct ggatatcctg gagtacatcc
     1261 atgagcatga gtacgtgcac ggggacatca aggcctccaa cctgctcctg agtcacaaga
     1321 accctgacca ggtatatttg gtagactatg gccttgctta tcggtactgc ccagatggag
     1381 ttcataaaga gtacaaggaa gatcccaaaa ggtgccatga cggcaccctg gagttcacca
     1441 gcatcgacgc tcacaaaggc gtggccccat caagacgtgg tgatttggaa atacttggtt
     1501 attgcatgat ccagtggctc agcggctgtc ttccttggga agataacttg aaagatccta
     1561 actacgttag ggattccaaa attagataca gagacaacgt cgcagctttg atggagaaat
     1621 gctttcctga gaaaaataag ccaggtgaga tcgctaagta catggagtct gtgaaactac
     1681 tggaatacac cgaaaaacct ctctatcaaa acctacgtga tatcctttta caaggactaa
     1741 aagctatagg aagtaaagac gacggcaaac tggattttag tgctgtggag aacggaagtg
     1801 tgaagacaag accagcctca aagaagcgga agaaagaagc agaagaaagc gcggtgtgcg
     1861 ctgtggagga catggagtgc tcagacacac aggtgcaaga ggccgcacag acccgaagtg
     1921 tggagtccca aggagcaata catggaagca tgtctcagcc agcggcgggc tgcagcagct
     1981 ctgacagttc caggaggcag cagcatttgg gattggaaca agacatgcta aggctagacc
     2041 gtaggggttc acgaaccaga aagaaagccc agaagggatc cgactacaaa gaccatgacg
     2101 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     2161 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2221 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2281 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2341 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2401 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2461 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2521 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     2581 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2641 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2701 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     2761 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     2821 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     2881 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     2941 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3001 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     3061 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3121 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3181 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3241 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3301 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3361 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3421 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     3481 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3541 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3601 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3721 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3781 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3841 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3901 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3961 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4021 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4081 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4141 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4201 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4261 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4321 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4381 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4441 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4501 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4561 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4621 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4681 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     4741 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4801 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4861 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4921 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4981 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5041 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5101 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5161 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5221 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5281 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5341 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5401 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5461 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5521 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5581 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5641 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5701 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5761 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5821 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5881 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5941 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6001 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6061 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6121 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6181 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6241 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6301 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6361 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6421 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6481 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6541 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6601 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6661 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6721 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6781 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6841 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6901 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6961 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7021 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7081 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7141 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7201 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     7261 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7321 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7381 aagaatctgc ttagg
//
