LOCUS       V031151                 8637 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031151
VERSION     V031151
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..4775
                     /label="Apaf1(NM_001042558)"
                     /note="Apaf1(NM_001042558)"
                     /gene="Apaf1"
     CDS             4803..4832
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             4851..4874
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(4894..4910)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    4921..5543
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(5692..6280)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    6609..6656
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(6888..7682)
                     /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(7717..8074)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(8076..8180)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      8207..8635
                     /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 ggatgcaaag gcccgcaatt gtttgcttca acatagagaa gctttggaaa
     1081 aggacatcaa aacatcctac atcatggatc acatgatcag taatggcgtc ttgtcagtga
     1141 tagaggagga gaaggtcaaa agtcaggcca ctcaatatca acgagcagcc gctttaatta
     1201 aaatgatact taataaagac aactgtgcct acatttcatt ctacaacgct ctgctacacg
     1261 agggctataa ggaccttgct gcgcttctgc agagtggcct tcctcttgtg tcgtcttcca
     1321 gtggcaagga cacagatggt ggaataactt catttgtaag gacagtgctg tgtgaaggtg
     1381 gagtacccca gaggccggtt attttcgtta ctagaaagaa gctggttcat gcgattcagc
     1441 agaagctctg gaaactgaat ggagaaccag ggtgggtcac catctatggg atggcaggct
     1501 gcggcaagtc tgtgttagct gcggaagccg ttcgagatca ctccctctta gaaggttgct
     1561 tttcaggggg tgtacactgg gtttccattg gaaaacaaga caaatctggg cttctcatga
     1621 aactgcagaa tctgtgcatg cgcttggacc aagaagagag tttctctcag aggcttccac
     1681 ttaatattga ggaggccaaa gaccgcctcc gtgttctgat gctgcgcaaa cacccaaggt
     1741 ctctgttgat cttggatgat gtttgggatc cttgggtgtt aaaagctttt gacaatcagt
     1801 gtcagattct tcttacaacc agagataaga gtgttacaga ttcagtaatg ggtcctaagc
     1861 atgttgtccc tgtggagagt ggtctaggga gagagaaagg acttgagatc ttgtcacttt
     1921 ttgttaacat gaagaaagaa gatctgccag cggaggctca cagtattata aaggaatgca
     1981 aaggttctcc tcttgtagtg tctttaattg gtgcgctttt acgtgatttt cccaatcgct
     2041 gggcatacta cctcagacag cttcagaata agcagtttaa gagaataagg aagtcttcat
     2101 cttatgatta tgaggctcta gatgaagcca tgtcgataag tgttgaaatg ctcagagaag
     2161 acatcaaaga ctattacaca gacctttcca tccttcagaa ggacgtcaag gtacctacaa
     2221 aggtgttgtg cgttctctgg gacttggaaa cggaagaagt tgaagacatc ctgcaggagt
     2281 tcgttaataa gtctctctta ttctgtaatc ggaatggaaa gtcattttgt tattatttac
     2341 atgatcttca agtagatttt cttacagaga agaatcgcag tcagcttcag gatctgcaca
     2401 ggaagatggt cactcagttt cagaggtatt accagcccca cacgctgtct ccagaccagg
     2461 aggactgcat gtattggtac aacttcctag cctatcacat ggctagtgcc aatatgcaca
     2521 aagaactttg tgctttaatg ttttccctgg actggattaa agcaaaaaca gaacttgtcg
     2581 gccctgccca tctgattcac gagttcgtgg catataggca tatattggat gaaaaggatt
     2641 gtgcagtctg tgagaatttt caagagtttt tatctttaaa tggacacctc cttggacgac
     2701 agccatttcc taatattgta cagctgggcc tctgtgaacc agaaacttcc gaagtttatc
     2761 gacaagcaaa gctgcaggcc aagcaggagg gggatactgg gcgcctttac ctggaatgga
     2821 taaacaaaaa aactatcaag aatctgtccc gcttagtcgt ccgcccccac acagatgctg
     2881 tttaccacgc gtgtttttct caggatggtc agagaatagc ttcttgtggg gctgataaaa
     2941 ccttacaggt gttcaaagcc gagacaggag agaaacttct tgacatcaaa gctcacgaag
     3001 atgaggtgct ttgctgcgcg ttctcctcag acgacagtta catagcgacc tgctcagcgg
     3061 ataagaaggt taagatttgg gattctgcga ctgggaagct tgtgcacacc tacgacgagc
     3121 actcggagca agtcaattgc tgccatttca ccaacaagag taaccacctt ctcttggcca
     3181 ctgggtcaaa tgatttcttc ctcaagctct gggatttgaa tcaaaaagaa tgtcgaaata
     3241 ccatgtttgg tcacacgaac tcagtcaacc actgcaggtt ctcaccagac gatgagctct
     3301 tggctagctg ctcagctgac gggactttaa ggctttggga tgtgagatca gcaaacgaga
     3361 ggaaaagcat taatgtgaag cgcttcttcc tgagttcaga agaccctcca gaggatgtgg
     3421 aggtgatcgt gaagtgttgt tcctggtctg cagatggtga caaaataata gtggcagcaa
     3481 aaaacaaagt cctccttttt gatattcata ctagtggcct attggcagag atccacacag
     3541 gccatcacag caccatccag tactgtgact tctcccccta tgaccatttg gctgtgattg
     3601 ccctgtctca gtactgtgtg gagttgtgga acatagactc ccgcctaaag gtggccgact
     3661 gcagaggaca tttgagttgg gttcacggtg tgatgttttc tcccgatggc tcctcatttt
     3721 tgacagcttc tgatgaccaa acaataaggg tctgggagac aaaaaaggta tgcaagaact
     3781 ctgccatcgt gctaaagcag gaaatagacg tcgtgtttca agagaacgaa acgatggtcc
     3841 ttgcagttga caacataaga ggcctgcaac tcattgctgg aaaaacaggc cagattgatt
     3901 acctgcctga agcccaagtg agttgctgct gcctcagtcc acaccttgag tacgtggcat
     3961 tcggagatga agatggagcc attaagatta tagaacttcc aaacaacaga gtcttcagtt
     4021 ctggggttgg gcacaagaaa gctgtgcggc acatccagtt cacagctgat gggaagacac
     4081 tgatttcaag ttctgaagat tctgtgattc aggtatggaa ttggcagaca ggggactatg
     4141 tatttttgca agcccaccag gaaacggtaa aggacttcag gctcctccaa gattcaagat
     4201 tgctttcttg gtcatttgat ggaacggtga aggtgtggaa tgtcattacc ggaagaatag
     4261 aaagagactt tacttgtcat cagggcacag tgctttcctg tgctatctct tctgatgcga
     4321 ccaagttttc atctacctct gctgataaga ctgccaagat ctggagtttt gacctccttt
     4381 cccctcttca tgagctgaag ggccataatg gctgtgtccg ctgctctgcc ttctcgctgg
     4441 atggcatcct gttggcaact ggagatgaca atggagaaat ccggatatgg aatgtctcag
     4501 atggccagct tcttcattcg tgtgctccga tctcggtaga ggaaggaact gctacccacg
     4561 gcggctgggt aactgatgtg tgcttctctc ccgacagtaa aacgcttgtc tctgctggag
     4621 gatatctcaa gtggtggaat gttgccactg gggactcctc acagaccttc tacacaaatg
     4681 gaacaaacct caagaaaatc cacgtgtccc ctgacttcag aacctatgtg actgtcgata
     4741 atctcggtat tttatatatt ttacaggttt tagagagcgg accgacgcgt acgcggccgc
     4801 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg
     4861 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc
     4921 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     4981 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     5041 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     5101 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     5161 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     5221 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     5281 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     5341 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     5401 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     5461 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc
     5521 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc
     5581 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5881 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6121 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6421 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     6481 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     6541 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     6601 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     6661 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     6721 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     6781 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     6841 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     6901 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     6961 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     7021 atgtcctgat agcgatccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     7081 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     7141 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     7201 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     7261 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     7321 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     7381 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     7441 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     7501 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     7561 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     7621 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     7681 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     7741 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     7801 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     7861 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     7921 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     7981 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     8041 acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca atattattga
     8101 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8161 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     8221 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     8281 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     8341 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8401 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     8461 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     8521 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     8581 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaatttt
//
