LOCUS       V026429                 7752 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026429
VERSION     V026429
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..1624
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1655..3241
                     /label="Runx2(NM_001278483)"
                     /note="Runx2(NM_001278483)"
                     /gene="Runx2"
     polyA_signal    3319..3543
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3589..4017
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4031..4360
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4427..5221
                     /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    5395..5516
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5565..5581)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5589..5605
                     /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(5613..5643)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5658..5679
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5967..6552)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6723..7583)
                     /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(7584..7688)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1081 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1321 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1381 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1501 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1621 caagggcggt ggaggcagtg gtggaggcgg tagcatggcg tcaaacagcc tcttcagtgc
     1681 agtgacaccg tgtcagcaaa acttcttttg ggatccgagc accagccggc gcttcagccc
     1741 cccctccagc agcctgcagc ccggcaagat gagcgacgtg agcccggtag tggctgctca
     1801 gcagcagcag cagcagcagc agcagcagca gcagcagcaa cagcagcaac agcagcagca
     1861 gcagcagcaa cagcagcagc agcagcagca ggaggcggcc gcagcagcag cagcggcagc
     1921 ggcggcggca gcagcggcgg cagccgtgcc ccggttgagg ccgccgcacg acaaccgcac
     1981 catggtggag atcatcgcgg accacccggc cgaactggtc cgcaccgaca gccccaactt
     2041 cctgtgctcc gtgctgccct cgcactggcg gtgcaacaag accctgcccg tggccttcaa
     2101 ggttgtagcc ctcggagagg tacccgatgg gaccgtggtt accgtcatgg ccgggaatga
     2161 tgagaactac tctgccgagc tacgaaatgc ctctgctgtt atgaaaaacc aagtggccag
     2221 gttcaacgat ctgagatttg taggccggag cggacgaggc aagagtttca ctttgaccat
     2281 aacggtcttc acaaatcctc cccaagtggc cacttaccac agagctatta aagtgacagt
     2341 ggacggtccc cgggaaccaa gaaggcacag acagaagctt gatgactcta aacctagttt
     2401 gttctctgac cgcctcagtg atttagggcg cattcctcat cccagtatga gagtaggtgt
     2461 cccgcctcag aacccacggc cctccctgaa ctcagcacca agtcctttta atccacaagg
     2521 acagagtcag attacagatc ccaggcagac acaatcctcc ccaccgtggt cctatgacca
     2581 gtcttacccc tcttatctga gccagatgac gtccccatcc atccattcca ccacgccgct
     2641 gtcttccaca cggggcactg ggctgccagc catcaccgac gtacccaggc gtatttcaga
     2701 tgatgacact gccacctctg acttctgcct ctggccttcc tctctcagta agaagagcca
     2761 ggcaggtgct tcagaactgg gccctttttc agaccccagg cagttcccag gcatttcatc
     2821 cctcactgag agccgcttct ccaacccacg aatgcactat ccagccacct tcacttacac
     2881 cccgccagtc acgtcaggca tgtccctggg catgtccgcc accactcact accacacgta
     2941 cctgccacca ccctaccccg gctcttccca aagccagagc ggacccttcc agaccagcag
     3001 cactccatat ctctactatg gtacttcgtc agcgtcctat cagttcccaa tggtccccgg
     3061 gggagaccgg tctccttcca ggatggtccc accgtgcacg accacctcga atggcagcac
     3121 gctattaaat ccaaatttgc ctaaccagaa tgatggtgtt gacgctgatg gaagccacag
     3181 cagctcccca accgttttga attctagcgg cagaatggat gagtctgtct ggcgacccta
     3241 ttgaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa
     3301 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3361 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3421 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3481 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3541 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3601 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3661 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3721 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3781 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3841 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3901 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3961 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4021 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4261 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4381 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4441 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4501 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4561 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4621 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4681 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4741 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4801 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4861 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4921 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4981 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5041 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5101 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5161 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     5221 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5281 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5341 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5401 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5461 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5521 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5581 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5641 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5701 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5761 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5821 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5881 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5941 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6001 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6061 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6121 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6181 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6241 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6301 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6361 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6421 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6481 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6601 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6661 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6721 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6781 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6841 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6901 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6961 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7021 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7081 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7141 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7201 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7261 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7321 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7381 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7441 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7501 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7561 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7621 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7681 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7741 atcccctatg gt
//
