LOCUS       V022981                 8477 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022981
VERSION     V022981
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3967
                     /label="PTK2B(NM_173176)"
                     /note="PTK2B(NM_173176)"
                     /gene="PTK2B"
     CDS             3974..4039
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4077..4301
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4347..4775
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4789..5118
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5185..5979
                     /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    6153..6274
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6323..6339)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6347..6363
                     /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(6371..6401)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6416..6437
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6725..7310)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7481..8341)
                     /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(8342..8446)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgtctgggg tgtccgagcc
      961 cctgagtcga gtaaagttgg gcacgttacg ccggcctgaa ggccctgcag agcccatggt
     1021 ggtggtacca gtagatgtgg aaaaggagga cgtgcgtatc ctcaaggtct gcttctatag
     1081 caacagcttc aatcctggga aaaacttcaa actggtcaaa tgcactgtcc agacggagat
     1141 ccgggagatc atcacctcca tcctgctgag cgggcggatc gggcccaaca tccggttggc
     1201 tgagtgctat gggctgaggc tgaagcacat gaagtccgat gagatccact ggctgcaccc
     1261 acagatgacg gtgggtgagg tgcaggacaa gtatgagtgt ctgcacgtgg aagccgagtg
     1321 gaggtatgac cttcaaatcc gctacttgcc agaagacttc atggagagcc tgaaggagga
     1381 caggaccacg ctgctctatt tttaccaaca gctccggaac gactacatgc agcgctacgc
     1441 cagcaaggtc agcgagggca tggccctgca gctgggctgc ctggagctca ggcggttctt
     1501 caaggatatg ccccacaatg cacttgacaa gaagtccaac ttcgagctcc tagaaaagga
     1561 agtggggctg gacttgtttt tcccaaagca gatgcaggag aacttaaagc ccaaacagtt
     1621 ccggaagatg atccagcaga ccttccagca gtacgcctcg ctcagggagg aggagtgcgt
     1681 catgaagttc ttcaacactc tcgccggctt cgccaacatc gaccaggaga cctaccgctg
     1741 tgaactcatt caaggatgga acattactgt ggacctggtc attggcccta aagggatccg
     1801 ccagctgact agtcaggacg caaagcccac ctgcctggcc gagttcaagc agatcaggtc
     1861 catcaggtgc ctcccgctgg aggagggcca ggcagtactt cagctgggca ttgaaggtgc
     1921 cccccaggcc ttgtccatca aaacctcatc cctagcagag gctgagaaca tggctgacct
     1981 catagacggc tactgccggc tgcagggtga gcaccaaggc tctctcatca tccatcctag
     2041 gaaagatggt gagaagcgga acagcctgcc ccagatcccc atgctaaacc tggaggcccg
     2101 gcggtcccac ctctcagaga gctgcagcat agagtcagac atctacgcag agattcccga
     2161 cgaaaccctg cgaaggcccg gaggtccaca gtatggcatt gcccgtgaag atgtggtcct
     2221 gaatcgtatt cttggggaag gcttttttgg ggaggtctat gaaggtgtct acacaaatca
     2281 caaaggggag aaaatcaatg tagctgtcaa gacctgcaag aaagactgca ctctggacaa
     2341 caaggagaag ttcatgagcg aggcagtgat catgaagaac ctcgaccacc cgcacatcgt
     2401 gaagctgatc ggcatcattg aagaggagcc cacctggatc atcatggaat tgtatcccta
     2461 tggggagctg ggccactacc tggagcggaa caagaactcc ctgaaggtgc tcaccctcgt
     2521 gctgtactca ctgcagatat gcaaagccat ggcctacctg gagagcatca actgcgtgca
     2581 cagggacatt gctgtccgga acatcctggt ggcctcccct gagtgtgtga agctggggga
     2641 ctttggtctt tcccggtaca ttgaggacga ggactattac aaagcctctg tgactcgtct
     2701 ccccatcaaa tggatgtccc cagagtccat taacttccga cgcttcacga cagccagtga
     2761 cgtctggatg ttcgccgtgt gcatgtggga gatcctgagc tttgggaagc agcccttctt
     2821 ctggctggag aacaaggatg tcatcggggt gctggagaaa ggagaccggc tgcccaagcc
     2881 tgatctctgt ccaccggtcc tttataccct catgacccgc tgctgggact acgaccccag
     2941 tgaccggccc cgcttcaccg agctggtgtg cagcctcagt gacgtttatc agatggagaa
     3001 ggacattgcc atggagcaag agaggaatgc tcgctaccga acccccaaaa tcttggagcc
     3061 cacagccttc caggaacccc cacccaagcc cagccgacct aagtacagac cccctccgca
     3121 aaccaacctc ctggctccaa agctgcagtt ccaggttcct gagggtctgt gtgccagctc
     3181 tcctacgctc accagcccta tggagtatcc atctcccgtt aactcactgc acaccccacc
     3241 tctccaccgg cacaatgtct tcaaacgcca cagcatgcgg gaggaggact tcatccaacc
     3301 cagcagccga gaagaggccc agcagctgtg ggaggctgaa aaggtcaaaa tgcggcaaat
     3361 cctggacaaa cagcagaagc agatggtgga ggactaccag tggctcaggc aggaggagaa
     3421 gtccctggac cccatggttt atatgaatga taagtcccca ttgacgccag agaaggaggt
     3481 cggctacctg gagttcacag ggcccccaca gaagcccccg aggctgggcg cacagtccat
     3541 ccagcccaca gctaacctgg accggactga tgacctggtg tacctcaatg tcatggagct
     3601 ggtgcgggcc gtgctggagc tcaagaatga gctctgtcag ctgccccccg agggctacgt
     3661 ggtggtggtg aagaatgtgg ggctgaccct gcggaagctc atcgggagcg tggatgatct
     3721 cctgccttcc ttgccgtcat cttcacggac agagatcgag ggcacccaga aactgctcaa
     3781 caaagacctg gcagagctca tcaacaagat gcggctggca cagcagaacg ccgtgacctc
     3841 cctaagtgag gagtgcaaga ggcagatgct gacggcttca cacaccctgg ctgtggacgc
     3901 caagaacctg ctcgacgctg tggaccaggc caaggttctg gccaatctgg cccacccacc
     3961 tgcagagctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     4021 caaggatgac gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt
     4081 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4141 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4201 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4261 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4321 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4381 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4441 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4501 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4561 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4621 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4681 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4741 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4801 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4861 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4921 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4981 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5041 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5101 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5161 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5221 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5281 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5341 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5401 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5461 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5521 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5581 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5641 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5701 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5761 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5821 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5881 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5941 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     6001 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6061 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6121 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     6181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6301 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6361 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6421 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6481 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6541 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6901 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7141 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8461 aagtgccacc tgacgtc
//
