LOCUS       sequence-9424-ex        9244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9244)
  AUTHORS   Liu B, Wang S, Brenner M, Paton JF, Kasparov S
  TITLE     Enhancement of cell-specific transgene expression from a Tet-Off 
            regulatory system using a transcriptional amplification strategy in 
            the rat brain.
  JOURNAL   J Gene Med. 2008 May . 10(5):583-92.
  PUBMED    18324737
REFERENCE   2  (bases 1 to 9244)
  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: "J Gene Med.
            2008 May . 10(5):583-92."
COMMENT     Sequence Label: pTYF-1xGfaABC1D-tTA
FEATURES             Location/Qualifiers
     source          1..9244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(838..1458,1459..1845)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(911..930)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    2072..2296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2429..2448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3469..3486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3640..4228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3720..3739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4399..5190,5191..5259))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5260..5364)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5432..5450
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5501..5523)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5609..5747
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5662..5681
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6951..6971
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    8592..8825
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 ggcctaggga taacagggta atagatcttg ctagccttgc atgcctgcag gtcggagtac
       61 tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt
      121 cctccgagcg gagactctag agggacgcgt aacatatcct ggtgtggagt aggggacgct
      181 gctctgacag aggctcgggg gcctgagctg gctctgtgag ctggggagga ggcagacagc
      241 caggccttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      301 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      361 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      421 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      481 ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc
      541 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      601 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      661 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
      721 gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc
      781 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggatccatg
      841 tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
      901 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
      961 tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat
     1021 aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat
     1081 aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta
     1141 ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc
     1201 caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact
     1261 ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct
     1321 actactgata gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt
     1381 gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt
     1441 aaatgtgaaa gtgggtccgc gtacagccgc gcgcgtacga aaaacaatta cgggtctacc
     1501 atcgagggcc tgctcgatct cccggacgac gacgcccccg aagaggcggg gctggcggct
     1561 ccgcgcctgt cctttctccc cgcgggacac acgcgcagac tgtcgacggc ccccccgacc
     1621 gatgtcagcc tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc
     1681 gacgcgctag acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga
     1741 tttacccccc acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag
     1801 cagatgttta ccgatgccct tggaattgac gagtacggtg ggtagctcga gcatgcatct
     1861 agatgatcag tcgactaggg ataacagggt aatggtacct ttaagaccaa tgacttacaa
     1921 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     1981 ctcccagggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     2041 aacccactgc ttaagcctca ataaagctcg actgtgcctt ctagttgcca gccatctgtt
     2101 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2161 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2221 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2281 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2341 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2461 acgttcgccg gctgtgcggg agaacggagt tctattatga ctcaaatcag tctccccaag
     2521 cattcgggga tcagagtttt taaggataac ttagtgtgta gggggccagt gagttggaga
     2581 tgaaagcgta gggagtcgaa ggtgtccttt tgcgccgagt cagttcctgg gtgggggcca
     2641 caagatcgga tgagccagtt tatcaatccg ggggtgccag ctgatccatg gagtgcaggg
     2701 tctgcaaaat atctcaagca ctgattgatc ttaggtttta caatagtgat gttaccccag
     2761 gaacaatttg gggaaggtca gaatcttgta gcctgtagct gcatgactcc taaaccataa
     2821 tttctttttt gttttttttt ttttattttt gagacagggt ctcactctgt cacctaggct
     2881 ggagtgcagt ggtgcaatca cagctcactg cagcctcaac gtcgtaagct caagcgatcc
     2941 tcccacctca gcctgcctgg tagctgagac tacaagcgac gccccagtta atttttgtat
     3001 ttttggtaga ggcagcgttt tgccgtgtgg ccctggctgg tctcgaactc ctgggctcaa
     3061 gtgatccagc ctcagcctcc caaagtgctg ggacaaccgg ggccagtcac tgcacctggc
     3121 cctaaaccat aatttctaat cttttggcta atttgttagt cctacaaagg cagtctagtc
     3181 cccaggcaaa aagggggttt gtttcgggaa agggctgtta ctgtctttgt ttcaaactat
     3241 aaactaagtt cctcctaaac ttagttcggc ctacacccag gaatgaacaa ggagagcttg
     3301 gaggttagaa gcacgatgga attggttagg tcagatctct ttcactgtct gagttataat
     3361 tttgcaatgg tggttcaaag actgcccgct tctgacacca gtcgctgcat taatgaatcg
     3421 gccaacgcgc ggggagaggc ggtttgcgta ttggcgctct tccgcttcct cgctcactga
     3481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     3841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4081 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4261 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4321 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4381 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4441 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4501 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4561 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4621 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4681 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4741 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4801 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4861 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4921 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4981 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5041 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5101 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5161 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5221 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5281 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5341 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5401 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5461 tcaagaactg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     5521 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     5581 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     5641 gagtgtactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5701 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     5761 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5821 gcggggaggc agagattgca gtaagctgag atcgcagcac tgcactccag cctgggcgac
     5881 agagtaagac tctgtctcaa aaataaaata aataaatcaa tcagatattc caatcttttc
     5941 ctttatttat ttatttattt tctattttgg aaacacagtc cttccttatt ccagaattac
     6001 acatatattc tatttttctt tatatgctcc agtttttttt agaccttcac ctgaaatgtg
     6061 tgtatacaaa atctaggcca gtccagcaga gcctaaaggt aaaaaataaa ataataaaaa
     6121 ataaataaaa tctagctcac tccttcacat caaaatggag atacagctgt tagcattaaa
     6181 taccaaataa cccatcttgt cctcaataat tttaagcgcc tctctccacc acatctaact
     6241 cctgtcaaag gcatgtgccc cttccgggcg ctctgctgtg ctgccaacca actggcatgt
     6301 ggactctgca gggtccctaa ctgccaagcc ccacagtgtg ccctgaggct gccccttcct
     6361 tctagcggct gcccccactc ggctttgctt tccctagttt cagttacttg cgttcagcca
     6421 aggtctgaaa ctaggtgcgc acagagcggt aagactgcga gagaaagaga ccagctttac
     6481 agggggttta tcacagtgca ccctgacagt cgtcagcctc acagggggtt tatcacattg
     6541 caccctgaca gtcgtcagcc tcacaggggg tttatcacag tgcaccctta caatcattcc
     6601 atttgattca caattttttt agtctctact gtgcctaact tgtaagttaa atttgatcag
     6661 aggtgtgttc ccagagggga aaacagtata tacagggttc agtactatcg catttcaggg
     6721 tggcccgaga gctgcatccg gagtatctag atggagttcc gcgttacata acttacggta
     6781 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgactgat
     6841 gttcccatag taacgccaat agggactttc cattgacgct aatgggagtt tgttttggca
     6901 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     6961 cggtaggcgt gtactctaga aggtctatat aagcagagct cgtttagtga accgtttata
     7021 ctacttatct ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     7081 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgctcaaagt agtgtgtgcc
     7141 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     7201 atctctagca gtggcgcccg aacagggact tgaaagcgaa agtaaagcca gaggagatct
     7261 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     7321 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     7381 tcggtattaa gcgggggaga attagataaa tgggaaaaaa ttcggttaag gccaggggga
     7441 aagaaacaat ataaactaaa acatatagta tgggcaagca gggagctaga acgattcgca
     7501 gttaatcctg gccttttaga gacatcagaa ggctgtagac aaatactggg acagctacaa
     7561 ccatcccttc agacaggatc agaagaactt agatcattat ataatacaat agcagtcctc
     7621 tattgtgtgc atcaaaggat agatgtaaaa gacaccaagg aagccttaga taagatagag
     7681 gaagagcaaa acaaaagtaa gaaaaaggca cagcaagcag cagctgacac aggaaacaac
     7741 agccaggtca gccaaaatta ccctatagtg cagaacctcc aggggcaaat ggtacatcag
     7801 gccatatcac ctagaacttt aaatgcatgg gtaaaagtag tagaagagaa ggctttcagc
     7861 ccagaagtaa tacccatgtt ttcagcatta tcagaaggag ccaccccaca agatttaaat
     7921 accatgctaa acacagtggg gggacatcaa gcagccatgc aaatgttaaa agagaccatc
     7981 aatgaggaag ctgcagaatg ggatagattg catccagtgc atgcagggcc tattgcacca
     8041 ggccagatga gagaaccaag gggaagtgac atagcaggaa ctactagcaa attaagagaa
     8101 caatttggaa ataataaaac aataatcttt aagcaatcct caggagggga cccagaaatt
     8161 gtaacgcaca gttttaattg tggaggggaa tttttctact gtaattcaac acaactgttt
     8221 aatagtactt ggtttaatag tacttggagt actgaagggt caaataacac tgaaggaagt
     8281 gacacaatca cactcccatg cagaataaaa caatttataa acatgtggca ggaagtagga
     8341 aaagcaatgt atgcccctcc catcagtgga caaattagat gttcatcaaa tattactggg
     8401 ctgctattaa caagagatgg tggtaataac aacaatgggt ccgagatctt cagacctgga
     8461 ggaggcgata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     8521 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     8581 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     8641 gcacggtcaa tgacgctgac ggtacaggcc agacaattat tgtctgatat agtgcagcag
     8701 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     8761 ggcatcaaac agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     8821 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     8881 gctagttgga gtaataaatc tctggaacag atttggaata acatgacctg gatggagtgg
     8941 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     9001 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcttg cggccggaat
     9061 tcacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     9121 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     9181 caaattacta aaaattcnaa aattttcggg tttattacag ggacagcaga gatccacttt
     9241 gggc
//
