LOCUS       V018852                 9155 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018852
VERSION     V018852
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             683..3802
                     /label="TNK2(NM_001308046)"
                     /note="TNK2(NM_001308046)"
                     /gene="TNK2"
     CDS             3842..3907
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3931..4519
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4585..4633
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4835..5263
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5277..5606
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5673..6467
                     /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    6643..6764
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6813..6829)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6837..6853
                     /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(6861..6891)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6906..6927
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7215..7803)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7974..8834)
                     /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(8835..8939)
                     /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 taccgagctc ggatccgcca ccatgccggc agcacgtcgg ttccctggcc tggagctctc
      721 cttccctctc ctggccagac tgcggcgacg cctgtacaca aggctgggag gcggcagaat
      781 gcagccagag gagggcacag gctggctgct ggagctgctg tccgaggtgc agctgcaaca
      841 gtacttcctg cggctccgag acgacctcaa cgtcacccgc ctgtcccact ttgagtacgt
      901 caagaatgag gacctggaga agatcggcat gggtcggcct ggccagcggc ggctgtggga
      961 ggctgtgaag aggaggaagg ccttgtgcaa acgcaagtcg tggatgagta aggtgttcag
     1021 tggaaagcga ctggaggctg agttcccacc tcatcactct cagagcacct tccggaagac
     1081 ctcgcccgcc cctgggggcc cagcagggga ggggcccctg cagagcctca cctgcctcat
     1141 tggggagaag gacctgcgcc tcctggagaa gctgggtgat ggttcctttg gcgtggtgcg
     1201 caggggcgag tgggacgcgc cctcagggaa gacggtgagt gtggctgtga agtgcctgaa
     1261 gcccgatgtc ctgagccagc cagaagccat ggacgacttc atccgggagg tcaatgccat
     1321 gcactcgctc gaccaccgaa acctcatccg cctctacggg gtggtgctca cgccgcccat
     1381 gaagatggtg acagagctgg cacctctggg atcgttgttg gaccggctac gtaagcacca
     1441 gggccacttc ctcctgggga ctctgagccg ctacgctgtg caggtggctg agggcatggg
     1501 ctacctggag tccaagcgct ttattcaccg tgacctggct gcccgcaatc tgctgttggc
     1561 tacccgcgac ctggtcaaga tcggggactt tgggctgatg cgagcactac ctcagaatga
     1621 cgaccattac gtcatgcagg aacatcgcaa ggtgcccttc gcctggtgtg cccccgagag
     1681 cctgaagaca cgtaccttct cccatgccag cgacacctgg atgttcgggg tgacactgtg
     1741 ggaaatgttc acctacggcc aggagccctg gatcggcctc aacggcagtc agatcctgca
     1801 taagatcgac aaggaggggg agcggctgcc ccggcccgag gactgtcccc aggacatcta
     1861 caacgtcatg gtccagtgct gggctcacaa gccagaggac agacccacgt ttgtggccct
     1921 gcgggacttc ctgctggagg cccagcccac agacatgcgg gcccttcagg actttgagga
     1981 accggacaag ctgcacatcc agatgaatga tgtcatcacc gtcatcgagg gaagggccga
     2041 gaactactgg tggcgtggcc agaacacacg gacgctgtgt gtggggccct tccctcgcaa
     2101 cgtggtgacc tccgtggccg gcctgtcggc ccaggacatc agccagcccc tgcagaacag
     2161 cttcatccac acagggcatg gcgacagtga cccccgccac tgctggggct tcccggacag
     2221 gattgacgaa ctgtatctgg gaaaccccat ggaccccccc gacctcctga gcgtggaact
     2281 gagcacctcc cggccccccc agcatctagg aggggtgaaa aaaccaacct atgaccctgt
     2341 gagcgaggac caagacccct tgtccagcga cttcaagagg ctgggcctgc ggaagccagg
     2401 cctgccccga gggctgtggc tggcgaagcc ctcggcgcgg gtgccgggca ccaaggccag
     2461 ccgaggcagc ggggctgagg tcacgctcat cgacttcggt gaggagcccg tggtcccggc
     2521 cctacggccc tgcgcgccct ccctggcgca gctggccatg gacgcctgct ccctgctgga
     2581 cgagaccccg cctcagagcc ccacgcgggc actgccccgg cccctgcacc ccacgcctgt
     2641 ggtggactgg gacgcacgcc cgctgccccc cccgcccgcc tatgacgacg tggcccagga
     2701 tgaggatgac tttgagatct gctccatcaa cagcaccctc gtgggcgcgg gggtccctgc
     2761 cgggcccagc cagggccaga ccaactacgc ctttgtgcct gagcaggcgc ggccgccccc
     2821 tcccctggag gacaacctgt tcctcccgcc ccagggtggg ggcaagccgc ccagctccgc
     2881 acagaccgca gagatcttcc aggcgctaca gcaggagtgc atgaggcaac tgcaggctcc
     2941 ggccggctcc ccggccccct ctcccagccc ggggggtgac gacaagcccc aggtgcctcc
     3001 tcgggtaccc atcccccctc ggcccacgcg cccacacgtc cagctgtctc cagccccccc
     3061 gggcgaggag gagaccagcc agtggcctgg acctgcttcc cctccccggg tgcctccgcg
     3121 ggagcccctg tcccctcaag gctcgaggac acccagcccc ctggtaccac ctggcagctc
     3181 cccgctgcca ccccggctct caagctcacc tgggaagacc atgcccacca cccagagctt
     3241 tgcctcagac cccaagtacg ccacccccca ggtgatccag gcccctggcc cgcgggctgg
     3301 tccctgcatc ctgcccatcg tccgggatgg caagaaggtc agcagcaccc actattactt
     3361 gctgcccgag cgaccatcct acctggagcg ctaccagcgc ttcctgcgtg aggcccagag
     3421 ccccgaggag cctacccccc tgcctgtgcc tctgctgctg cccccaccca gcaccccagc
     3481 ccccgccgcc cccacggcca ccgtgcggcc gatgccccag gctgccttgg accccaaggc
     3541 caacttctcc accaacaaca gcaacccagg ggcccggcca ccacccccga gggccactgc
     3601 tcggctgcca cagaggggct gccctggcga tgggccagag gcgggccggc cagcagacaa
     3661 gatccagatg gtggagcagc tcttcgggct gggtctgcgg cccagagggg agtgccacaa
     3721 agtgctggag atgttcgact ggaacctgga gcaggccggc tgccaccttc tgggctcctg
     3781 gggccctgcc caccacaagc gcgaattctg cagatatcca gcacagtggc ggccgctcga
     3841 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3901 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3961 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4021 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4081 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4141 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4201 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4261 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4321 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4381 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4441 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4501 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     4561 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     4621 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4681 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4741 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     4801 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4861 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4921 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4981 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5041 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5101 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5161 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     5221 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5281 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5341 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5401 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5521 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5581 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5641 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5701 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5761 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5821 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5881 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5941 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     6001 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     6061 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6121 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6181 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6241 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6301 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6361 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6421 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6481 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6541 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6601 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6661 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6721 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6781 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6841 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6901 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6961 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7021 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7081 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7141 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7201 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7261 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7321 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7381 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7441 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7501 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7561 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7621 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7681 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7741 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7801 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7861 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7921 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7981 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8041 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8101 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8161 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8221 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8281 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8341 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8401 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8461 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8521 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8581 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8641 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8701 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8761 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8821 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8881 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8941 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     9001 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     9061 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     9121 gcttgaccga caattgcatg aagaatctgc ttagg
//
