LOCUS       V031239                 6423 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031239
VERSION     V031239
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..2570
                     /label="Runx2(NM_001145920)"
                     /note="Runx2(NM_001145920)"
                     /gene="Runx2"
     CDS             2589..2618
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2637..2660
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     primer_bind     complement(2680..2696)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     polyA_signal    2707..3329
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     rep_origin      complement(3478..4066)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     polyA_signal    4395..4442
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     CDS             complement(4674..5468)
                     /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(5503..5860)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        complement(5862..5966)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      5993..6421
                     /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 gcgtattcct gtagatccga gcaccagccg gcgcttcagc cccccctcca
     1081 gcagcctgca gcccggcaag atgagcgacg tgagcccggt ggtggctgcg cagcagcagc
     1141 aacagcagca gcagcagcag caacagcagc agcaacaaca gcaacagcaa caacagcagc
     1201 agcagcagca gcagcaggag gcggccgcag cagcagcggc ggcagcggcg gcggcagcag
     1261 cggcggcggc cgcagtgccc cgattgaggc cgccgcacga caaccgcacc atggtggaga
     1321 tcatcgcgga ccacccggcc gaactggtcc gcaccgacag tcccaacttc ctgtgctccg
     1381 tgctgccctc gcactggcgg tgcaacaaga ccctgcccgt ggccttcaag gttgtagccc
     1441 tcggagaggt accagatggg actgtggtta ccgtcatggc cgggaatgat gagaactact
     1501 ccgccgagct ccgaaatgcc tccgctgtta tgaaaaacca agtagccagg ttcaacgatc
     1561 tgagatttgt gggccggagc ggacgaggca agagtttcac cttgaccata acagtcttca
     1621 caaatcctcc ccaagtggcc acttaccaca gagctattaa agtgacagtg gacggtcccc
     1681 gggaaccaag aaggcacaga cagaagcttg atgactctaa acctagtttg ttctctgatc
     1741 gcctcagtga tttagggcgc attcctcatc ccagtatgag agtaggtgtc ccgcctcaga
     1801 acccacggcc ctccctgaac tctgcaccaa gtccttttaa tccacaagga cagagtcaga
     1861 ttacagatcc caggcaggca cagtcttccc caccgtggtc ctatgaccag tcttacccct
     1921 cctatctgag ccagatgaca tccccatcca tccactccac cacgccgctg tcttccacac
     1981 ggggcaccgg gctacctgcc atcactgacg tgcccaggcg tatttcagat gatgacactg
     2041 ccacctctga cttctgcctc tggccttcct ctctcagtaa gaagagccag gcaggtgctt
     2101 cagaactggg ccctttttca gaccccaggc agttcccaag catttcatcc ctcactgaga
     2161 gccgcttctc caacccacga atgcactacc cagccacctt tacctacacc ccgccagtca
     2221 cgtcaggcat gtccctcggc atgtccgcca ccactcacta ccacacgtac ctgccaccac
     2281 cctaccccgg ctcttcccaa agccagagtg gacccttcca gaccagcagc actccatatc
     2341 tctactatgg tacttcgtca gcatcctatc agttcccaat ggtacccggg ggagaccggt
     2401 ctccttccag gatggtccca ccatgcacca ccacctcgaa tggcagcacg ctattaaatc
     2461 caaatttgcc taaccagaat gatggtgttg acgctgacgg aagccacagc agttccccaa
     2521 ctgttttgaa ttctagcggc agaatggatg agtctgtttg gcggccatat acgcgtacgc
     2581 ggccgctcga gcagaaactc atctcagaag aggatctggc agcaaatgat atcctggatt
     2641 acaaggatga cgacgataag gtttaaacgg ccggccgcgg tcatagctgt ttcctgaaca
     2701 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     2761 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2821 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     2881 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     2941 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     3001 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     3061 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     3121 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     3181 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     3241 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     3301 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcggcgag cggtatcagc
     3361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3901 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4201 aatctaaagt atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc
     4261 tgcgagccct gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg
     4321 ggcgtattgg ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     4381 aaccccgcgt ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc
     4441 cgtcatagcg cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct
     4501 agggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
     4561 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
     4621 tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag
     4681 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     4741 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     4801 aacgctatgt cctgatagcg atccgccaca cccagccggc cacagtcgat gaatccagaa
     4861 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     4921 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     4981 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     5041 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     5101 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     5161 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     5221 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     5281 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     5341 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     5401 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     5461 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc
     5521 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     5581 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     5641 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     5701 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     5761 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     5821 taactgacac acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat
     5881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc
     6001 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6061 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6121 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6181 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6241 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6301 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6361 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6421 ttt
//
