LOCUS       sequence-271971-        7324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible mammalian co-expression vector for human bone 
            morphogenetic protein 2 and 7 cDNAs (2 transcriptional units, 
            divergent).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7324)
  AUTHORS   Feichtinger GA, Hacobian A, Hofmann AT, Wassermann K, Zimmermann A, 
            van Griensven M, Redl H
  TITLE     Constitutive and inducible co-expression systems for non-viral 
            osteoinductive gene therapy.
  JOURNAL   Eur Cell Mater. 2014 Feb 19;27:166-84; discussion 184.
  PUBMED    24554273
REFERENCE   2  (bases 1 to 7324)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eur Cell 
            Mater."; date: "2014-02-19"; volume: "27"; pages: "166-84; 
            discussion 184"
COMMENT     Sequence Label: pTetON-BMP2/7
FEATURES             Location/Qualifiers
     source          1..7324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    71..89
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    107..125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    143..161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    178..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    214..232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      377..386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      381..390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1750..1831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1842..1859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2291..2425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2382..2401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2435..2443,2444..2479,2480..2482,2483..2518,
                     2519..2521,2522..2557,2558..2563,2564..3181))
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(3284..3487)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3488..3867
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3948..4536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4028..4047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4707..5498,5499..5567))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5330..5349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5568..5672)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5705..5786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        join(7243..7324,1..306)
                     /label=bidirectional TRE promoter
                     /note="Tet-responsive bidirectional promoter PTight-BI, 
                     consisting of seven tet operator sequences flanked on each 
                     side by the minimal CMV promoter"
     protein_bind    join(7324,1..18)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
       61 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      121 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      181 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      241 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      301 atcgcctgga gatttcgagc tcggtaccga gctcggatcc actagtaacg gccgccagtg
      361 tgctggaatt cggcttgcca ccatggtggc cgggacccgc tgtcttctag cgttgctgct
      421 tccccaggtc ctcctgggcg gcgcggctgg cctcgttccg gagctgggcc gcaggaagtt
      481 cgcggcggcg tcgtcgggcc gcccctcatc ccagccctct gacgaggtcc tgagcgagtt
      541 cgagctgcgg ctgctcagca tgttcggcct gaaacagaga cccaccccca gcagggacgc
      601 cgtggtgccc ccctacatgc tagacctgta tcgcaggcac tcaggtcagc cgggctcacc
      661 cgccccagac caccggttgg agagggcagc cagccgagcc aacactgtgc gcagcttcca
      721 ccatgaagaa tctttggaag aactaccaga aacgagtggg aaaacaaccc ggagattctt
      781 ctttaattta agttctatcc ccacggagga gtttatcacc tcagcagagc ttcaggtttt
      841 ccgagaacag atgcaagatg ctttaggaaa caatagcagt ttccatcacc gaattaatat
      901 ttatgaaatc ataaaacctg caacagccaa ctcgaaattc cccgtgacca gacttttgga
      961 caccaggttg gtgaatcaga atgcaagcag gtgggaaagt cttgatgtca cccccgctgt
     1021 gatgcggtgg actgcacagg gacacgccaa ccatggattc gtggtggaag tggcccactt
     1081 ggaggagaaa caaggtgtct ccaagagaca tgttaggata agcaggtctt tgcaccaaga
     1141 tgaacacagc tggtcacaga taaggccatt gctagtaact tttggccatg atggaaaagg
     1201 gcatcctctc cacaaaagag aaaaacgtca agccaaacac aaacagcgga aacgccttaa
     1261 gtccagctgt aagagacacc ctttgtacgt ggacttcagt gacgtggggt ggaatgactg
     1321 gattgtggct cccccggggt atcacgcctt ttactgccac ggagaatgcc cttttcctct
     1381 ggctgatcat ctgaactcca ctaatcatgc cattgttcag acgttggtca actctgttaa
     1441 ctctaagatt cctaaggcat gctgtgtccc gacagaactc agtgctatct cgatgctgta
     1501 ccttgacgag aatgaaaagg ttgtattaaa gaactatcag gacatggttg tggagggttg
     1561 tgggtgtcgc tagaagccga attctgcaga tatccatcac actggcggcc gcatcgataa
     1621 gcttgtcgac gatatctcca gaggatcata atcagccata ccacatttgt agaggtttta
     1681 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1741 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1801 aatttcacaa ataaagcatt tttttcactg ccccgagctt cctcgctcac tgactcgctg
     1861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1921 tccacagaat caggggataa cgcaggaaag aacatgtttg gtcgagctga tacttcccgt
     1981 ccgccagggg gacatgccgg cgatgctgaa ggtcgcgcgc attcccgatg aagaggccgg
     2041 ttaccgcctg ttgacctggt gggacgggca gggcgccgcc cgagtcttcg cctcggcggc
     2101 gggcgctctg ctcatggagc gcgcgtccgg ggccggggac cttgcacaga tagcgtggtc
     2161 cggccaggac gacgaggctt gcaggatcat aatcagccat accacatttg tagaggtttt
     2221 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2281 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2341 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2401 caatgtatct tatcatgtct ggatccttac ttagttaccc ggggagcatg tcaaggtcaa
     2461 aatcgtcaag agcgtcagca ggcagcatat caaggtcaaa gtcgtcaagg gcatcggctg
     2521 ggagcatgtc taagtcaaaa tcgtcaaggg cgtcggccgg cccgccgctt tcgcacttta
     2581 gctgtttctc caggccacat atgattagtt ccaggccgaa aaggaaggca ggttcggctc
     2641 cctgccggtc gaacagctca attgcttgtc tcagaagtgg gggcatagaa tcggtggtag
     2701 gtgtctctct ttcctctttt gctacttgat gctcctgttc ctccaatacg cagcccagtg
     2761 taaagtggcc cacggcggac agagcgtaca gtgcgttctc cagggagaag ccttgctgac
     2821 acaggaacgc gagctgattt tccagggttt cgtactgttt ctctgttggg cgggtgccga
     2881 gatgcacttt agccccgtcg cgatgtgaga ggagagcaca gcggaatgac ttggcgttgt
     2941 tccgcagaaa gtcttgccat gactcgcctt ccagggggca gaagtgggta tgatgcctgt
     3001 ccagcatctc gattggcagg gcatcgagca gggcccgctt gttcttcacg tgccagtaca
     3061 gggtaggctg ctcaactccc agcttttgag cgagtttcct tgtcgtcagg ccttcgatac
     3121 cgactccatt gagtaattcc agagcgccgt ttatgacttt gctcttgtcc agtctagaca
     3181 tggtgaattc ggggccgcgg aggctggatc ggtcccggtg tcttctatgg aggtcaaaac
     3241 agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg cttatataga
     3301 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     3361 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     3421 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     3481 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3541 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     3601 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     3661 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     3721 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3781 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     3841 attaataact agtcaataat caatgtcaac atggcggtaa tgttggacat gtgagcaaaa
     3901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     5341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5701 gtcggcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     5761 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     5821 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     5881 ctgattatga tcctctagac tgcagcctca ggagatctct agtggcagcc acaggcccgg
     5941 accaccatgt ttctgtattt cttcaggatg acgttggagc tgtcatcgaa gtagaggacg
     6001 gagatggcat tgagctgcgt gggcgcacag cagggcttgg gcaccgtttc cgggttgatg
     6061 aagtggacca gcgtctgcac gatggcgtgg ttggtggcgt tcatgtagga gttcagaggg
     6121 aaggcacact ccccctcaca gtagtaggcg gcgtagcctt caggcgcgat gatccagtcc
     6181 tgccagccca ggtctcggaa gctgacatac agctcgtgct tcttacaggc ctgcctctgg
     6241 tcgctgctgc tgttctctgc cacgttggcc atccgcaggg cttcctggtt cttgggcgtc
     6301 ttggagcggt tctggctgcg ctgtttgctc cccgtggacc ggatgctgcg gaagtggacc
     6361 tccgtggcct tgaagaaagc caccatgaag ggctgcttgt tctggggccc gtgccgccca
     6421 atcaggcccg ccaacttggg gttgatgctc tgcccatcca gcgtctccac cgagagctgc
     6481 aggcccaggt tgtgccgcgg attgaccacc cagtggttgc tggtggctgt gatgtcaaac
     6541 accagccagc cctcctccga ggcccagagg gtacggctgt cgagcaggaa gagatccgat
     6601 tccctgccca agtgctcctg gagcacctga taaacgctga tccggaacgt ctcattgtcg
     6661 aagcgttccc ggatgtagtc cttgtagatc cggaattcgg ctgccgtgac agcttcccct
     6721 tctgggatct tggaaagatc aaaccggaac tctcgatggt ggtagcgtgg gtggaagaat
     6781 tccttgtcat gttccacgag gttgacgaag ctcatgacca tgtcggcgtc ggtgaggaaa
     6841 tggctatctt gcaggctggc cagagggggg ccctgggtac tgaagacggc cttgtagggg
     6901 taggagaagc cctggccgcc gggcccgccg ccctcctcca ccgccatggc gttgtacagg
     6961 tccagcatga acatgggtgc cgagttgtgc ttgccctgga ggtgcgggcg cgggcggtgg
     7021 ggcaagccca aaatggagag gatctcgcgc tgcatctccc gccgctcctg gctgcggagg
     7081 cgccggtgga tgaagctcga gtgcacctcg ttgtccaggc tgaagtcggc cagggcggag
     7141 cgcagcagga acaggggtgc ccagagcgcc acgaagctgt gcggcgccgc agctcgcagt
     7201 gagcgcacgt gcatggtggc catatgaccg gtgaattctc caggcgatct gacggttcac
     7261 taaacgagct ctgcttatat aggcctccca ccgtacacgc cacctcgaca tactcgagtt
     7321 tact
//
