LOCUS       sequence-9426-ex        9084 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 9084)
  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 9084)
  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-1xSYN-tTA
FEATURES             Location/Qualifiers
     source          1..9084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(678..1298,1299..1685)
                     /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(751..770)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    1912..2136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2269..2288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3309..3326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4239..5030,5031..5099))
                     /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"
     primer_bind     4862..4881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5100..5204)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5272..5290
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5341..5363)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5449..5587
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5502..5521
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6791..6811
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    8432..8665
                     /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 atcgatggcg ccagctgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgactc tagagctgca ggcaagcttg
      661 gtaccgagct cggatccatg tctagattag ataaaagtaa agtgattaac agcgcattag
      721 agctgcttaa tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag
      781 gtgtagagca gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct
      841 tagccattga gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct
      901 ggcaagattt tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg
      961 atggagcaaa agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa
     1021 atcaattagc ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca
     1081 gcgctgtggg gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta
     1141 aagaagaaag ggaaacacct actactgata gtatgccgcc attattacga caagctatcg
     1201 aattatttga tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat
     1261 gcggattaga aaaacaactt aaatgtgaaa gtgggtccgc gtacagccgc gcgcgtacga
     1321 aaaacaatta cgggtctacc atcgagggcc tgctcgatct cccggacgac gacgcccccg
     1381 aagaggcggg gctggcggct ccgcgcctgt cctttctccc cgcgggacac acgcgcagac
     1441 tgtcgacggc ccccccgacc gatgtcagcc tgggggacga gctccactta gacggcgagg
     1501 acgtggcgat ggcgcatgcc gacgcgctag acgatttcga tctggacatg ttgggggacg
     1561 gggattcccc gggtccggga tttacccccc acgactccgc cccctacggc gctctggata
     1621 tggccgactt cgagtttgag cagatgttta ccgatgccct tggaattgac gagtacggtg
     1681 ggtagctcga gcatgcatct agatgatcag tcgactaggg ataacagggt aatggtacct
     1741 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     1801 ggactggaag ggctaattca ctcccagggt ctctctggtt agaccagatc tgagcctggg
     1861 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagctcg actgtgcctt
     1921 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1981 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2041 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2101 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2161 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2221 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2281 tcttcccttc ctttctcgcc acgttcgccg gctgtgcggg agaacggagt tctattatga
     2341 ctcaaatcag tctccccaag cattcgggga tcagagtttt taaggataac ttagtgtgta
     2401 gggggccagt gagttggaga tgaaagcgta gggagtcgaa ggtgtccttt tgcgccgagt
     2461 cagttcctgg gtgggggcca caagatcgga tgagccagtt tatcaatccg ggggtgccag
     2521 ctgatccatg gagtgcaggg tctgcaaaat atctcaagca ctgattgatc ttaggtttta
     2581 caatagtgat gttaccccag gaacaatttg gggaaggtca gaatcttgta gcctgtagct
     2641 gcatgactcc taaaccataa tttctttttt gttttttttt ttttattttt gagacagggt
     2701 ctcactctgt cacctaggct ggagtgcagt ggtgcaatca cagctcactg cagcctcaac
     2761 gtcgtaagct caagcgatcc tcccacctca gcctgcctgg tagctgagac tacaagcgac
     2821 gccccagtta atttttgtat ttttggtaga ggcagcgttt tgccgtgtgg ccctggctgg
     2881 tctcgaactc ctgggctcaa gtgatccagc ctcagcctcc caaagtgctg ggacaaccgg
     2941 ggccagtcac tgcacctggc cctaaaccat aatttctaat cttttggcta atttgttagt
     3001 cctacaaagg cagtctagtc cccaggcaaa aagggggttt gtttcgggaa agggctgtta
     3061 ctgtctttgt ttcaaactat aaactaagtt cctcctaaac ttagttcggc ctacacccag
     3121 gaatgaacaa ggagagcttg gaggttagaa gcacgatgga attggttagg tcagatctct
     3181 ttcactgtct gagttataat tttgcaatgg tggttcaaag actgcccgct tctgacacca
     3241 gtcgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggcgctct
     3301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4201 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4261 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4321 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4381 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4441 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4501 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4561 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4621 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4681 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4741 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4801 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4861 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4921 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4981 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5041 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5221 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5281 atcacgaggc cctttcgtct tcaagaactg cctcgcgcgt ttcggtgatg acggtgaaaa
     5341 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5401 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     5461 ccagtcacgt agcgatagcg gagtgtactg gcttaactat gcggcatcag agcagattgt
     5521 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5581 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5641 ctcttcgcta ttacgccagc gcggggaggc agagattgca gtaagctgag atcgcagcac
     5701 tgcactccag cctgggcgac agagtaagac tctgtctcaa aaataaaata aataaatcaa
     5761 tcagatattc caatcttttc ctttatttat ttatttattt tctattttgg aaacacagtc
     5821 cttccttatt ccagaattac acatatattc tatttttctt tatatgctcc agtttttttt
     5881 agaccttcac ctgaaatgtg tgtatacaaa atctaggcca gtccagcaga gcctaaaggt
     5941 aaaaaataaa ataataaaaa ataaataaaa tctagctcac tccttcacat caaaatggag
     6001 atacagctgt tagcattaaa taccaaataa cccatcttgt cctcaataat tttaagcgcc
     6061 tctctccacc acatctaact cctgtcaaag gcatgtgccc cttccgggcg ctctgctgtg
     6121 ctgccaacca actggcatgt ggactctgca gggtccctaa ctgccaagcc ccacagtgtg
     6181 ccctgaggct gccccttcct tctagcggct gcccccactc ggctttgctt tccctagttt
     6241 cagttacttg cgttcagcca aggtctgaaa ctaggtgcgc acagagcggt aagactgcga
     6301 gagaaagaga ccagctttac agggggttta tcacagtgca ccctgacagt cgtcagcctc
     6361 acagggggtt tatcacattg caccctgaca gtcgtcagcc tcacaggggg tttatcacag
     6421 tgcaccctta caatcattcc atttgattca caattttttt agtctctact gtgcctaact
     6481 tgtaagttaa atttgatcag aggtgtgttc ccagagggga aaacagtata tacagggttc
     6541 agtactatcg catttcaggg tggcccgaga gctgcatccg gagtatctag atggagttcc
     6601 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6661 tgacgtcaat aatgactgat gttcccatag taacgccaat agggactttc cattgacgct
     6721 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc
     6781 gccccgttga cgcaaatggg cggtaggcgt gtactctaga aggtctatat aagcagagct
     6841 cgtttagtga accgtttata ctacttatct ggtctctctg gttagaccag atctgagcct
     6901 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6961 tgctcaaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7021 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     7081 agtaaagcca gaggagatct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     7141 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     7201 gagagatggg tgcgagagcg tcggtattaa gcgggggaga attagataaa tgggaaaaaa
     7261 ttcggttaag gccaggggga aagaaacaat ataaactaaa acatatagta tgggcaagca
     7321 gggagctaga acgattcgca gttaatcctg gccttttaga gacatcagaa ggctgtagac
     7381 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     7441 ataatacaat agcagtcctc tattgtgtgc atcaaaggat agatgtaaaa gacaccaagg
     7501 aagccttaga taagatagag gaagagcaaa acaaaagtaa gaaaaaggca cagcaagcag
     7561 cagctgacac aggaaacaac agccaggtca gccaaaatta ccctatagtg cagaacctcc
     7621 aggggcaaat ggtacatcag gccatatcac ctagaacttt aaatgcatgg gtaaaagtag
     7681 tagaagagaa ggctttcagc ccagaagtaa tacccatgtt ttcagcatta tcagaaggag
     7741 ccaccccaca agatttaaat accatgctaa acacagtggg gggacatcaa gcagccatgc
     7801 aaatgttaaa agagaccatc aatgaggaag ctgcagaatg ggatagattg catccagtgc
     7861 atgcagggcc tattgcacca ggccagatga gagaaccaag gggaagtgac atagcaggaa
     7921 ctactagcaa attaagagaa caatttggaa ataataaaac aataatcttt aagcaatcct
     7981 caggagggga cccagaaatt gtaacgcaca gttttaattg tggaggggaa tttttctact
     8041 gtaattcaac acaactgttt aatagtactt ggtttaatag tacttggagt actgaagggt
     8101 caaataacac tgaaggaagt gacacaatca cactcccatg cagaataaaa caatttataa
     8161 acatgtggca ggaagtagga aaagcaatgt atgcccctcc catcagtgga caaattagat
     8221 gttcatcaaa tattactggg ctgctattaa caagagatgg tggtaataac aacaatgggt
     8281 ccgagatctt cagacctgga ggaggcgata tgagggacaa ttggagaagt gaattatata
     8341 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     8401 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     8461 cagcaggaag cactatgggc gcacggtcaa tgacgctgac ggtacaggcc agacaattat
     8521 tgtctgatat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     8581 tgttgcaact cacagtctgg ggcatcaaac agctccaggc aagaatcctg gctgtggaaa
     8641 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     8701 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaata
     8761 acatgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     8821 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     8881 aatgggcttg cggccggaat tcacaaatgg cagtattcat ccacaatttt aaaagaaaag
     8941 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     9001 aaactaaaga attacaaaaa caaattacta aaaattcnaa aattttcggg tttattacag
     9061 ggacagcaga gatccacttt gggc
//
