LOCUS       V019210                 7589 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019210
VERSION     V019210
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2269
                     /label="Runx2(NM_001278483)"
                     /note="Runx2(NM_001278483)"
                     /gene="Runx2"
     CDS             2276..2341
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2365..2953
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3019..3067
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3269..3697
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3711..4040
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4107..4901
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5077..5198
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5247..5263)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5271..5287
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5295..5325)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5340..5361
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5649..6237)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6408..7268)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7269..7373)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcgtc aaacagcctc ttcagtgcag tgacaccgtg
      721 tcagcaaaac ttcttttggg atccgagcac cagccggcgc ttcagccccc cctccagcag
      781 cctgcagccc ggcaagatga gcgacgtgag cccggtagtg gctgctcagc agcagcagca
      841 gcagcagcag cagcagcagc agcagcaaca gcagcaacag cagcagcagc agcagcaaca
      901 gcagcagcag cagcagcagg aggcggccgc agcagcagca gcggcagcgg cggcggcagc
      961 agcggcggca gccgtgcccc ggttgaggcc gccgcacgac aaccgcacca tggtggagat
     1021 catcgcggac cacccggccg aactggtccg caccgacagc cccaacttcc tgtgctccgt
     1081 gctgccctcg cactggcggt gcaacaagac cctgcccgtg gccttcaagg ttgtagccct
     1141 cggagaggta cccgatggga ccgtggttac cgtcatggcc gggaatgatg agaactactc
     1201 tgccgagcta cgaaatgcct ctgctgttat gaaaaaccaa gtggccaggt tcaacgatct
     1261 gagatttgta ggccggagcg gacgaggcaa gagtttcact ttgaccataa cggtcttcac
     1321 aaatcctccc caagtggcca cttaccacag agctattaaa gtgacagtgg acggtccccg
     1381 ggaaccaaga aggcacagac agaagcttga tgactctaaa cctagtttgt tctctgaccg
     1441 cctcagtgat ttagggcgca ttcctcatcc cagtatgaga gtaggtgtcc cgcctcagaa
     1501 cccacggccc tccctgaact cagcaccaag tccttttaat ccacaaggac agagtcagat
     1561 tacagatccc aggcagacac aatcctcccc accgtggtcc tatgaccagt cttacccctc
     1621 ttatctgagc cagatgacgt ccccatccat ccattccacc acgccgctgt cttccacacg
     1681 gggcactggg ctgccagcca tcaccgacgt acccaggcgt atttcagatg atgacactgc
     1741 cacctctgac ttctgcctct ggccttcctc tctcagtaag aagagccagg caggtgcttc
     1801 agaactgggc cctttttcag accccaggca gttcccaggc atttcatccc tcactgagag
     1861 ccgcttctcc aacccacgaa tgcactatcc agccaccttc acttacaccc cgccagtcac
     1921 gtcaggcatg tccctgggca tgtccgccac cactcactac cacacgtacc tgccaccacc
     1981 ctaccccggc tcttcccaaa gccagagcgg acccttccag accagcagca ctccatatct
     2041 ctactatggt acttcgtcag cgtcctatca gttcccaatg gtccccgggg gagaccggtc
     2101 tccttccagg atggtcccac cgtgcacgac cacctcgaat ggcagcacgc tattaaatcc
     2161 aaatttgcct aaccagaatg atggtgttga cgctgatgga agccacagca gctccccaac
     2221 cgttttgaat tctagcggca gaatggatga gtctgtctgg cgaccctatc tcgaggacta
     2281 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     2341 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     2401 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2461 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2521 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2581 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2641 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2701 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     2761 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2821 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2881 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2941 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     3001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     3061 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     3121 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     3181 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     3241 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3301 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3361 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3421 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3481 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3541 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3601 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     3661 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3721 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3901 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3961 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4021 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4081 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4141 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4201 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4261 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4321 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4381 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4441 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4501 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4561 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4621 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4681 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4741 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4801 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4861 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4921 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4981 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5041 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5101 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5161 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5221 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5281 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5341 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5401 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5461 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5521 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5581 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5641 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5701 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5761 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5821 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5881 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5941 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6001 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6061 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6361 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6421 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6481 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6541 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6601 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6661 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6721 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6781 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6841 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6901 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6961 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7021 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7081 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7141 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7201 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7261 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7321 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7381 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     7441 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7501 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7561 ccgacaattg catgaagaat ctgcttagg
//
