LOCUS       V021863                 6734 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021863
VERSION     V021863
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             1004..2896
                     /label="Trp73(NM_011642)"
                     /note="Trp73(NM_011642)"
                     /gene="Trp73"
     CDS             2900..2929
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2948..2971
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2991..3007)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    3018..3640
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3789..4377)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4706..4753
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4985..5779)
                     /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(5814..6171)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(6173..6277)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      6304..6732
                     /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 tggatccgcc accatggccc agacctcttc
     1021 ttcctcctcc tccaccttcg agcacctgtg gagttctcta gagccagaca gcacctactt
     1081 tgacctcccc cagcccagcc aagggactag cgaggcatca ggcagcgagg agtccaacat
     1141 ggatgtcttc cacctgcaag gcatggccca gttcaatttg ctcagcagtg ccatggacca
     1201 gatgggcagc cgtgcggccc cggcgagccc ctacaccccg gagcacgccg ccagcgcgcc
     1261 cacccactcg ccctacgcgc agcccagctc caccttcgac accatgtctc cggcgcctgt
     1321 catcccttcc aataccgact accccggccc ccaccacttc gaggtcacct tccagcagtc
     1381 gagcactgcc aagtcggcca cctggacata ctccccactc ttgaagaagt tgtactgtca
     1441 gattgctaag acatgcccca tccagatcaa agtgtccaca ccaccacccc cgggcacggc
     1501 catccgggcc atgcctgtct acaagaaggc agagcatgtg accgacattg ttaagcgctg
     1561 ccccaaccac gagcttggaa gggacttcaa tgaaggacag tctgccccgg ctagccacct
     1621 catccgtgta gaaggcaaca acctcgccca gtacgtggat gaccctgtca ccggaaggca
     1681 gagtgtggtt gtgccgtatg aacccccaca ggtgggaaca gaatttacca ccatcctgta
     1741 caacttcatg tgtaacagca gctgtgtggg gggcatgaat cggaggccca tccttgtcat
     1801 catcaccctg gagacccggg atggacaggt cctgggccgc cggtctttcg agggtcgcat
     1861 ctgtgcctgt cctggccgtg accgcaaagc tgatgaagac cattaccggg agcaacaggc
     1921 tctgaatgaa agtaccacca aaaatggagc tgccagcaaa cgtgcattca agcagagccc
     1981 ccctgccatc cctgccctgg gtaccaacgt gaagaagaga cgccacgggg acgaggacat
     2041 gttctacatg cacgtgcgag gccgggagaa ctttgagatc ttgatgaaag tcaaggagag
     2101 cctagaactg atggagcttg tgccccagcc tttggttgac tcctatcgac agcagcagca
     2161 gcagcagctc ctacagaggc cgagtcacct gcagcctcca tcctatgggc ccgtgctctc
     2221 cccaatgaac aaggtacacg gtggtgtcaa caaactgccc tccgtcaacc agctggtggg
     2281 ccagcctccc ccgcacagct cagcagctgg gcccaacctg gggcccatgg gctccgggat
     2341 gctcaacagc cacggccaca gcatgccggc caatggtgag atgaatggag gccacagctc
     2401 ccagaccatg gtttcgggat cccactgcac cccgccaccc ccctatcatg cagaccccag
     2461 cctcgtcagt tttttgacag ggttggggtg tccaaactgc atcgagtgct tcacttccca
     2521 agggttgcag agcatctacc acctgcagaa ccttaccatc gaggaccttg gggctctgaa
     2581 ggtccctgac cagtaccgta tgaccatctg gaggggccta caggacctga agcagagcca
     2641 tgactgcggc cagcaactgc tacgctccag cagcaacgcg gccaccatct ccatcggcgg
     2701 ctctggcgag ctgcagcggc agcgggtcat ggaagccgtg catttccgtg tgcgccacac
     2761 catcacgatc cccaaccgtg gaggcgcagg tgcggtgaca ggtcccgacg agtgggcgga
     2821 ctttggcttt gacctgcctg actgcaagtc ccgtaagcag cccatcaaag aggagttcac
     2881 agagacagag agccacctcg agcagaaact catctcagaa gaggatctgg cagcaaatga
     2941 tatcctggat tacaaggatg acgacgataa ggtttaaacg gccggccgcg gtcatagctg
     3001 tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     3061 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     3121 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     3181 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     3241 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     3301 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     3361 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     3421 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     3481 ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc cagacacagc
     3541 ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt ggcactgtcc
     3601 tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg ccagcggcga
     3661 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3721 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3781 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3841 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3901 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3961 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4021 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4081 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4141 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4201 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4261 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4321 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4381 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4441 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4501 agttttaaat caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc
     4561 ccgacgttgg ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag
     4621 gaagaaacgc gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag
     4681 ccctgggacc gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt
     4741 ctttttattg ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt
     4801 agcctccccc tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     4861 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     4921 tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     4981 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     5041 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     5101 cacgggtagc caacgctatg tcctgatagc gatccgccac acccagccgg ccacagtcga
     5161 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     5221 tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg
     5281 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     5341 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     5401 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     5461 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     5521 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     5581 gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     5641 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     5701 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     5761 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa
     5821 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     5881 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     5941 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     6001 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     6061 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     6121 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc
     6181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6301 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6361 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6421 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6481 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6541 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6601 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6661 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6721 ttaacgcgaa tttt
//
