LOCUS       V031284                 9018 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031284
VERSION     V031284
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..5165
                     /label="Met(NM_001410120)"
                     /note="Met(NM_001410120)"
                     /gene="Met"
     CDS             5184..5213
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             5232..5255
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(5275..5291)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    5302..5924
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(6073..6661)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6990..7037
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(7269..8063)
                     /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(8098..8455)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8457..8561)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8588..9016
                     /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 gaaggctccc accgtgctgg cacctggcat tctggtgctg ctgttgtcct
     1081 tggtgcagag gagccatggg gagtgcaagg aggccctagt gaagtctgag atgaacgtga
     1141 acatgaagta tcagctcccc aacttcacgg cagaaacccc catccagaat gtcgtcctac
     1201 acggccatca tatttatctc ggagccacaa actacattta tgttttaaat gacaaagacc
     1261 ttcagaaggt atccgaattc aagaccgggc ccgtgttgga acacccagat tgtttacctt
     1321 gtcgggactg cagcagcaaa gccaattcat caggaggggt ttggaaagac aacatcaaca
     1381 tggctctgct tgttgacaca tactatgatg atcaactcat tagctgtggc agtgtcaaca
     1441 gagggacttg ccagcggcat gtccttcctc ctgacaattc tgctgacatc cagtctgagg
     1501 tccactgcat gttctcccca gaagaggagt cagggcagtg tcctgactgt gtagtgagtg
     1561 ccctcggagc caaagtcctc ctgtcggaaa aggaccggtt catcaatttc tttgtgggga
     1621 atacgatcaa ttcctcctat cctcctggtt attcactgca ttcgatatcg gtgagacggc
     1681 tgaaggaaac ccaagatggt tttaagtttt tgacagacca gtcctatatt gatgtcttac
     1741 cagaattcca agattcctac cccataaagt acatacatgc cttcgaaagc aaccatttta
     1801 tttactttct gactgtccaa aaggaaactc tagatgctca gacttttcat acaagaataa
     1861 tcaggttctg ttccgtagac tctgggttgc actcctacat ggaaatgccc ctggaatgca
     1921 tcctgacaga aaaaagaagg aagagatcca caagggaaga agtgtttaat atcctccaag
     1981 ccgcgtatgt cagtaaacca ggggccaatc ttgctaagca aataggagct agcccttctg
     2041 atgacattct cttcggggtg tttgcacaaa gcaagccaga ttctgctgaa cctgtgaatc
     2101 gatcagcagt ctgtgcattc cccatcaaat atgtcaatga cttcttcaac aagattgtca
     2161 acaaaaacaa cgtgagatgt ctccagcatt tttacggacc caaccatgag cactgtttca
     2221 ataggaccct gctgagaaac tcttccggct gtgaagcgcg cagtgacgag tatcggacag
     2281 agtttaccac ggctttgcag cgcgtcgact tattcatggg ccggcttaac caagtgctcc
     2341 tgacatccat ctccaccttc atcaaaggtg acctcaccat tgctaatcta gggacgtcag
     2401 aaggtcgctt catgcaggtg gtgctctctc gaacagcaca cctcactcct catgtgaact
     2461 tcctcctgga ctcccatcct gtatctccag aagttattgt tgagcatcca tcaaatcaaa
     2521 atggctatac attggttgtc acaggaaaga agatcaccaa gattccattg aatggcctgg
     2581 gctgtggaca tttccaatcc tgcagtcagt gcctctctgc cccttacttt atacagtgtg
     2641 gctggtgcca caatcaatgt gtgcgttttg atgaatgccc cagcggtaca tggactcaag
     2701 agatctgtct gccggcggtt tataaggtgt tccccaccag cgcgcccctt gaaggaggaa
     2761 cagtgttgac catatgtggc tgggactttg gattcaggaa gaataataaa tttgatttaa
     2821 ggaaaaccaa agttctgctt ggcaacgaga gctgtacctt gaccttaagc gagagcacga
     2881 caaatacgtt gaaatgcaca gttggtcccg cgatgagtga gcacttcaat gtgtctgtaa
     2941 ttatctcaaa cagtcgagag acgacgcaat acagtgcatt ctcctatgta gatcctgtaa
     3001 taacaagcat ttctccgagg tacggccctc aggctggagg caccttactc actcttactg
     3061 ggaaatacct caacagtggc aattctagac acatttcaat tggagggaaa acatgtactt
     3121 taaaaagtgt atcagatagt attcttgaat gctacacccc agcccaaact acctctgatg
     3181 agtttcctgt gaaattgaag attgacttgg ctaaccgaga gaccagcagc ttcagttacc
     3241 gggaagaccc cgttgtctat gaaatccacc cgaccaaatc ttttattagt ggtggaagca
     3301 caataacggg tattgggaag accctgaact cggttagcct cccaaagctg gtaatagatg
     3361 tgcatgaagt gggtgtgaac tacacagtgg catgtcagca tcgctcaaat tcagagatca
     3421 tctgctgcac tactccttca ctgaaacagc tgggcctgca actccccctg aagaccaaag
     3481 ccttcttcct gttagacggg attctttcca aacactttga tctcacttat gtgcataatc
     3541 ctgtgtttga gccttttgaa aagccagtaa tgatctcaat aggcaatgaa aatgtagtgg
     3601 aaattaaggg aaacaatatt gaccctgaag cagttaaagg tgaagtgtta aaagttggaa
     3661 atcagagctg cgagagtctc cactggcact ctggagctgt gttgtgtaca gtccccagtg
     3721 acctgctcaa actgaacagc gagctaaata tagagtggaa gcaagcagtc tcttcaactg
     3781 ttcttggaaa agtgatcgtt caaccggatc agaattttgc aggattgatc attggtgcgg
     3841 tctcaatatc agtagtagtt ttgttattat ccgggctctt cctgtggatg agaaagagaa
     3901 agcataaaga tctgggcagt gaattagttc gctatgacgc aagagtacac actcctcatt
     3961 tggataggct tgtaagtgcc cgaagtgtaa gtccaactac agagatggtt tcaaatgagt
     4021 ctgtagacta cagagctact tttccagaag accagtttcc caactcctct cagaatggag
     4081 catgcagaca agtgcaatac cctctgacag acctgtcccc tatcctgaca agtggagact
     4141 ctgatatatc cagcccatta ctacaaaata ctgttcacat tgacctcagt gctctaaatc
     4201 cagagctggt ccaagcagtt cagcacgtag tgattggacc cagcagcctg attgtgcatt
     4261 tcaatgaagt cataggaaga gggcattttg gctgtgtcta tcatgggact ttgctggaca
     4321 atgacggaaa gaaaattcac tgtgctgtga aatccttaaa tagaatcaca gatatagaag
     4381 aggtctccca gtttctgact gagggaatca tcatgaaaga cttcagccat cccaatgttc
     4441 tctcactctt gggaatctgc ctgaggagtg aagggtctcc tctggtggtc ctgccctata
     4501 tgaagcatgg agatctgcga aatttcattc gaaacgagac tcataatcca actgtgaaag
     4561 atcttatagg atttggcctt caagtagcca aaggcatgaa atatcttgcc agcaaaaagt
     4621 ttgtccacag agacttagct gcaagaaact gcatgttgga tgaaaaattc actgtcaagg
     4681 ttgctgattt cggtcttgcc agagacatgt acgataaaga gtactatagt gtccacaaca
     4741 agacgggtgc caagctacca gtaaagtgga tggctttaga gagtctgcaa acgcagaagt
     4801 tcaccaccaa gtcagatgtg tggtcctttg gtgtgctcct ctgggagctc atgacgagag
     4861 gagcccctcc ttatcccgac gtgaacacat ttgatatcac tatctacctg ttgcaaggca
     4921 gaagactctt gcaaccagaa tactgtccag acgccttgta cgaagtgatg ctaaaatgct
     4981 ggcaccccaa agcggaaatg cgcccgtcct tttccgaact ggtctccagg atatcctcaa
     5041 tcttctccac gttcattggg gaacactacg tccacgtgaa cgctacttat gtgaatgtaa
     5101 aatgtgttgc tccatatcct tctctgttgc catcccaaga caacattgat ggcgagggga
     5161 acacaacgcg tacgcggccg ctcgagcaga aactcatctc agaagaggat ctggcagcaa
     5221 atgatatcct ggattacaag gatgacgacg ataaggttta aacggccggc cgcggtcata
     5281 gctgtttcct gaacagatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     5341 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     5401 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     5461 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     5521 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     5581 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     5641 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     5701 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     5761 gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca
     5821 cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact
     5881 gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagcg
     5941 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6001 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6061 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6121 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6181 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6241 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6301 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6361 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6421 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6481 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6541 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6601 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6661 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6721 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6781 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     6841 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     6901 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     6961 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     7021 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     7081 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     7141 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     7201 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     7261 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     7321 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     7381 atatcacggg tagccaacgc tatgtcctga tagcgatccg ccacacccag ccggccacag
     7441 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     7501 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     7561 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     7621 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     7681 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     7741 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     7801 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     7861 gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc ggtcttgaca
     7921 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     7981 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     8041 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcgatcttt
     8101 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     8161 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     8221 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     8281 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     8341 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     8401 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
     8461 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     8521 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8581 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     8641 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     8701 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     8761 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     8821 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     8881 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     8941 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     9001 aaatttaacg cgaatttt
//
