LOCUS       V030017                 6900 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030017
VERSION     V030017
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     166..182
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        343..722
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        723..926
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        952..970
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             1029..3047
                     /label="Prkcb(NM_008855)"
                     /note="Prkcb(NM_008855)"
                     /gene="Prkcb"
     CDS             3066..3095
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             3114..3137
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(3157..3173)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3184..3806
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3955..4543)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4872..4919
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(5151..5945)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     promoter        complement(5980..6337)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6339..6443)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6470..6898
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat ggctgacccg gctgcggggc cgccgccgag cgagggcgag gagagcacag
     1081 tgcgcttcgc ccgcaaaggc gccctccggc agaagaacgt gcacgaggtg aagaaccaca
     1141 aattcaccgc ccgcttcttc aagcagccca ccttctgcag ccactgcacc gacttcatct
     1201 ggggcttcgg gaagcaggga ttccagtgtc aagtctgctg ctttgttgta cacaagcgct
     1261 gccatgagtt cgtcacgttc tcctgccccg gtgcggacaa gggcccggcc tctgatgacc
     1321 cccggagcaa acacaagttt aagatccaca cgtactccag tcctaccttc tgtgaccact
     1381 gtggatcgct gctgtatgga cttattcacc aggggatgaa atgcgacacc tgtatgatga
     1441 acgtgcacaa gcgctgcgtg atgaacgtcc ccagcctctg cggcactgac cacacggaac
     1501 gccgcggccg catctatatc caggcccaca tcgacagaga ggttctcatt gttgttgtaa
     1561 gagatgctaa aaatctggta cctatggacc ccaatggctt gtcagatccc tacgttaaac
     1621 tgaaactgat ccctgatccc aaaagtgaga gcaaacagaa gaccaagacc atcaagtgtt
     1681 ccctcaatcc agagtggaat gaaaccttca gatttcagct gaaggaatca gacaaagaca
     1741 gaagactgtc tgtagagatc tgggattggg acctgaccag caggaatgac ttcatgggat
     1801 ctctgtcatt tgggatttca gaactacaga aagccggtgt ggatggctgg ttcaagttac
     1861 taagccagga agagggtgaa tacttcaatg tgcccgtgcc gcccgaagga agcgagggca
     1921 atgaagagct gcggcagaag tttgagagag ccaagattgg ccaaggtacc aaggctccag
     1981 aagaaaagac agctaacact atctccaaat ttgacaacaa tggcaacagg gaccggatga
     2041 aactgaccga ttttaacttc ctgatggtgc tggggaaagg cagctttggc aaggtcatgc
     2101 tctcagagcg gaagggtaca gatgaactct atgccgtgaa gatcctgaag aaggacgtgg
     2161 tgatccaaga tgacgatgtg gagtgcacca tggtggagaa gcgggtgctg gccctgcctg
     2221 ggaagccccc gttcctgact cagctccatt cctgcttcca gaccatggac cgcctgtact
     2281 ttgtgatgga gtatgtgaac gggggtgacc tcatgtacca catccaacaa gttggccgtt
     2341 tcaaggagcc ccatgctgta ttttacgccg cagagattgc catcggtctg ttcttcttgc
     2401 agagcaaggg cattatttac cgcgacctga aacttgacaa cgtgatgctg gattctgagg
     2461 ggcacatcaa aatcgctgac tttggcatgt gtaaggagaa catctgggat ggggtgacaa
     2521 ccaagacatt ctgtggcact ccggactaca ttgctccaga gatcattgct tatcagccct
     2581 acgggaagtc tgtggactgg tgggcatttg gagtcctgct gtatgaaatg ttggctggcc
     2641 aggcaccgtt tgaaggggag gatgaggatg aactcttcca gtcaatcatg gaacacaatg
     2701 tggcgtatcc caagtccatg tccaaggaag ccgtggcaat ctgcaaaggg ctaatgacca
     2761 aacacccagg caagcgcctg ggttgtgggc ctgaagggga acgagacatt aaggagcatg
     2821 cgtttttccg gtatattgac tgggagaaac tcgaacgcaa ggagattcag ccaccttata
     2881 aaccaaaagc ttgtgggcga aacgctgaaa acttcgaccg gtttttcacc cgccatccac
     2941 cagtcctaac acctcctgac caggaagtca tcaggaatat tgaccaatca gaattcgaag
     3001 gattttcctt tgttaactct gaatttttaa aacctgaagt caagagcacg cgtacgcggc
     3061 cgctcgagca gaaactcatc tcagaagagg atctggcagc aaatgatatc ctggattaca
     3121 aggatgacga cgataaggtt taaacggccg gccgcggtca tagctgtttc ctgaacagat
     3181 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     3241 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     3301 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     3361 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct
     3421 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag
     3481 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga
     3541 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca
     3601 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt
     3661 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc
     3721 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg
     3781 tccactcagt agatgcctgt tgaattgggt acgcggccag cggcgagcgg tatcagctca
     3841 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3901 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3961 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4021 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4081 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4141 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4201 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4261 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4321 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4381 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4441 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4501 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4561 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4621 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4681 ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc
     4741 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
     4801 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
     4861 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
     4921 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg
     4981 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     5041 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     5101 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     5161 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     5221 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     5281 gctatgtcct gatagcgatc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     5341 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     5401 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     5461 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     5521 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     5581 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     5641 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     5701 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     5761 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     5821 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     5881 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     5941 atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc
     6001 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     6061 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     6121 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     6181 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     6241 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     6301 ctgacacaca ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat
     6361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     6481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6541 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6661 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6781 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6841 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
//
