LOCUS       sequence-266308-        7305 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 7305)
  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 7305)
  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..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    84..102
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        185..223
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      318..327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      322..331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1691..1772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1783..1800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2232..2366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2323..2342
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2376..2384,2385..2420,2421..2423,2424..2459,
                     2460..2462,2463..2498,2499..2504,2505..3122))
                     /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(3225..3428)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3429..3808
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3889..4477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3969..3988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4648..5439,5440..5508))
                     /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     5271..5290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5509..5613)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5646..5727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    7265..7283
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
       61 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttatc
      121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
      181 tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca
      241 gatcgcctgg agatttcgag ctcggtaccg agctcggatc cactagtaac ggccgccagt
      301 gtgctggaat tcggcttgcc accatggtgg ccgggacccg ctgtcttcta gcgttgctgc
      361 ttccccaggt cctcctgggc ggcgcggctg gcctcgttcc ggagctgggc cgcaggaagt
      421 tcgcggcggc gtcgtcgggc cgcccctcat cccagccctc tgacgaggtc ctgagcgagt
      481 tcgagctgcg gctgctcagc atgttcggcc tgaaacagag acccaccccc agcagggacg
      541 ccgtggtgcc cccctacatg ctagacctgt atcgcaggca ctcaggtcag ccgggctcac
      601 ccgccccaga ccaccggttg gagagggcag ccagccgagc caacactgtg cgcagcttcc
      661 accatgaaga atctttggaa gaactaccag aaacgagtgg gaaaacaacc cggagattct
      721 tctttaattt aagttctatc cccacggagg agtttatcac ctcagcagag cttcaggttt
      781 tccgagaaca gatgcaagat gctttaggaa acaatagcag tttccatcac cgaattaata
      841 tttatgaaat cataaaacct gcaacagcca actcgaaatt ccccgtgacc agacttttgg
      901 acaccaggtt ggtgaatcag aatgcaagca ggtgggaaag tcttgatgtc acccccgctg
      961 tgatgcggtg gactgcacag ggacacgcca accatggatt cgtggtggaa gtggcccact
     1021 tggaggagaa acaaggtgtc tccaagagac atgttaggat aagcaggtct ttgcaccaag
     1081 atgaacacag ctggtcacag ataaggccat tgctagtaac ttttggccat gatggaaaag
     1141 ggcatcctct ccacaaaaga gaaaaacgtc aagccaaaca caaacagcgg aaacgcctta
     1201 agtccagctg taagagacac cctttgtacg tggacttcag tgacgtgggg tggaatgact
     1261 ggattgtggc tcccccgggg tatcacgcct tttactgcca cggagaatgc ccttttcctc
     1321 tggctgatca tctgaactcc actaatcatg ccattgttca gacgttggtc aactctgtta
     1381 actctaagat tcctaaggca tgctgtgtcc cgacagaact cagtgctatc tcgatgctgt
     1441 accttgacga gaatgaaaag gttgtattaa agaactatca ggacatggtt gtggagggtt
     1501 gtgggtgtcg ctagaagccg aattctgcag atatccatca cactggcggc cgcatcgata
     1561 agcttgtcga cgatatctcc agaggatcat aatcagccat accacatttg tagaggtttt
     1621 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1681 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1741 aaatttcaca aataaagcat ttttttcact gccccgagct tcctcgctca ctgactcgct
     1801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1861 atccacagaa tcaggggata acgcaggaaa gaacatgttt ggtcgagctg atacttcccg
     1921 tccgccaggg ggacatgccg gcgatgctga aggtcgcgcg cattcccgat gaagaggccg
     1981 gttaccgcct gttgacctgg tgggacgggc agggcgccgc ccgagtcttc gcctcggcgg
     2041 cgggcgctct gctcatggag cgcgcgtccg gggccgggga ccttgcacag atagcgtggt
     2101 ccggccagga cgacgaggct tgcaggatca taatcagcca taccacattt gtagaggttt
     2161 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2221 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2281 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2341 tcaatgtatc ttatcatgtc tggatcctta cttagttacc cggggagcat gtcaaggtca
     2401 aaatcgtcaa gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag ggcatcggct
     2461 gggagcatgt ctaagtcaaa atcgtcaagg gcgtcggccg gcccgccgct ttcgcacttt
     2521 agctgtttct ccaggccaca tatgattagt tccaggccga aaaggaaggc aggttcggct
     2581 ccctgccggt cgaacagctc aattgcttgt ctcagaagtg ggggcataga atcggtggta
     2641 ggtgtctctc tttcctcttt tgctacttga tgctcctgtt cctccaatac gcagcccagt
     2701 gtaaagtggc ccacggcgga cagagcgtac agtgcgttct ccagggagaa gccttgctga
     2761 cacaggaacg cgagctgatt ttccagggtt tcgtactgtt tctctgttgg gcgggtgccg
     2821 agatgcactt tagccccgtc gcgatgtgag aggagagcac agcggaatga cttggcgttg
     2881 ttccgcagaa agtcttgcca tgactcgcct tccagggggc agaagtgggt atgatgcctg
     2941 tccagcatct cgattggcag ggcatcgagc agggcccgct tgttcttcac gtgccagtac
     3001 agggtaggct gctcaactcc cagcttttga gcgagtttcc ttgtcgtcag gccttcgata
     3061 ccgactccat tgagtaattc cagagcgccg tttatgactt tgctcttgtc cagtctagac
     3121 atggtgaatt cggggccgcg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa
     3181 cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag
     3241 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     3301 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     3361 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     3421 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     3481 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     3541 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     3601 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     3661 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     3721 agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac
     3781 tattaataac tagtcaataa tcaatgtcaa catggcggta atgttggaca tgtgagcaaa
     3841 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3901 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3961 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4021 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4081 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4141 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4201 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4261 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4321 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4381 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4441 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4501 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4561 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4681 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4741 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4801 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4861 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4921 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4981 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5041 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5101 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5161 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5221 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     5281 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5341 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5401 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5461 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5521 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5581 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5641 cgtcggcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5701 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5761 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     5821 gctgattatg atcctctaga ctgcagcctc aggagatctc tagtggcagc cacaggcccg
     5881 gaccaccatg tttctgtatt tcttcaggat gacgttggag ctgtcatcga agtagaggac
     5941 ggagatggca ttgagctgcg tgggcgcaca gcagggcttg ggcaccgttt ccgggttgat
     6001 gaagtggacc agcgtctgca cgatggcgtg gttggtggcg ttcatgtagg agttcagagg
     6061 gaaggcacac tccccctcac agtagtaggc ggcgtagcct tcaggcgcga tgatccagtc
     6121 ctgccagccc aggtctcgga agctgacata cagctcgtgc ttcttacagg cctgcctctg
     6181 gtcgctgctg ctgttctctg ccacgttggc catccgcagg gcttcctggt tcttgggcgt
     6241 cttggagcgg ttctggctgc gctgtttgct ccccgtggac cggatgctgc ggaagtggac
     6301 ctccgtggcc ttgaagaaag ccaccatgaa gggctgcttg ttctggggcc cgtgccgccc
     6361 aatcaggccc gccaacttgg ggttgatgct ctgcccatcc agcgtctcca ccgagagctg
     6421 caggcccagg ttgtgccgcg gattgaccac ccagtggttg ctggtggctg tgatgtcaaa
     6481 caccagccag ccctcctccg aggcccagag ggtacggctg tcgagcagga agagatccga
     6541 ttccctgccc aagtgctcct ggagcacctg ataaacgctg atccggaacg tctcattgtc
     6601 gaagcgttcc cggatgtagt ccttgtagat ccggaattcg gctgccgtga cagcttcccc
     6661 ttctgggatc ttggaaagat caaaccggaa ctctcgatgg tggtagcgtg ggtggaagaa
     6721 ttccttgtca tgttccacga ggttgacgaa gctcatgacc atgtcggcgt cggtgaggaa
     6781 atggctatct tgcaggctgg ccagaggggg gccctgggta ctgaagacgg ccttgtaggg
     6841 gtaggagaag ccctggccgc cgggcccgcc gccctcctcc accgccatgg cgttgtacag
     6901 gtccagcatg aacatgggtg ccgagttgtg cttgccctgg aggtgcgggc gcgggcggtg
     6961 gggcaagccc aaaatggaga ggatctcgcg ctgcatctcc cgccgctcct ggctgcggag
     7021 gcgccggtgg atgaagctcg agtgcacctc gttgtccagg ctgaagtcgg ccagggcgga
     7081 gcgcagcagg aacaggggtg cccagagcgc cacgaagctg tgcggcgccg cagctcgcag
     7141 tgagcgcacg tgcatggtgg ccatatgacc ggtgaattct ccaggcgatc tgacggttca
     7201 ctaaacgagc tctgcttata taggcctccc accgtacacg ccacctcgac atactcgagt
     7261 ttactcccta tcagtgatag agaacgtatg tcgagtttac tccct
//
