LOCUS       sequence-9429-ex        8499 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 8499)
  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 8499)
  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-Tretight-DsRed2
FEATURES             Location/Qualifiers
     source          1..8499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..317
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    11..29
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    47..65
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    82..100
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    118..136
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    154..172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    225..243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      356..365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             362..1039
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(542..562)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     954..977
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1249..1473
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1606..1625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2646..2663)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2817..3405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2897..2916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3576..4367,4368..4436))
                     /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     4199..4218
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4437..4541)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4609..4627
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4678..4700)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4786..4924
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4839..4858
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    7769..8002
                     /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 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
       61 agagaacgat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc
      121 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      181 cgagtttatc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
      241 agaacgtatg tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag
      301 tgaaccgtca gatcgcctgg agaattcgag ctcggtaccc ggggatccac cggtcgccac
      361 catggcctcc tccgagaacg tcatcaccga gttcatgcgc ttcaaggtgc gcatggaggg
      421 caccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
      481 ccacaacacc gtgaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
      541 cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc
      601 cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
      661 ggacggcggc gtggcgaccg tgacccagga ctcctccctg caggacggct gcttcatcta
      721 caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtgatgc agaagaagac
      781 catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
      841 gacccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtccat
      901 ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggacg ccaagctgga
      961 catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgca ccgagggccg
     1021 ccaccacctg ttcctgtagc ggccgcatcg attagggata acagggtaat ggtaccttta
     1081 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     1141 ctggaagggc taattcactc ccagggtctc tctggttaga ccagatctga gcctgggagc
     1201 tctctggcta actagggaac ccactgctta agcctcaata aagctcgact gtgccttcta
     1261 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1321 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1381 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1441 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1501 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1561 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1621 tcccttcctt tctcgccacg ttcgccggct gtgcgggaga acggagttct attatgactc
     1681 aaatcagtct ccccaagcat tcggggatca gagtttttaa ggataactta gtgtgtaggg
     1741 ggccagtgag ttggagatga aagcgtaggg agtcgaaggt gtccttttgc gccgagtcag
     1801 ttcctgggtg ggggccacaa gatcggatga gccagtttat caatccgggg gtgccagctg
     1861 atccatggag tgcagggtct gcaaaatatc tcaagcactg attgatctta ggttttacaa
     1921 tagtgatgtt accccaggaa caatttgggg aaggtcagaa tcttgtagcc tgtagctgca
     1981 tgactcctaa accataattt cttttttgtt tttttttttt tatttttgag acagggtctc
     2041 actctgtcac ctaggctgga gtgcagtggt gcaatcacag ctcactgcag cctcaacgtc
     2101 gtaagctcaa gcgatcctcc cacctcagcc tgcctggtag ctgagactac aagcgacgcc
     2161 ccagttaatt tttgtatttt tggtagaggc agcgttttgc cgtgtggccc tggctggtct
     2221 cgaactcctg ggctcaagtg atccagcctc agcctcccaa agtgctggga caaccggggc
     2281 cagtcactgc acctggccct aaaccataat ttctaatctt ttggctaatt tgttagtcct
     2341 acaaaggcag tctagtcccc aggcaaaaag ggggtttgtt tcgggaaagg gctgttactg
     2401 tctttgtttc aaactataaa ctaagttcct cctaaactta gttcggccta cacccaggaa
     2461 tgaacaagga gagcttggag gttagaagca cgatggaatt ggttaggtca gatctctttc
     2521 actgtctgag ttataatttt gcaatggtgg ttcaaagact gcccgcttct gacaccagtc
     2581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg gcgctcttcc
     2641 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2701 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2761 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2821 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2881 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2941 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3001 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3061 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3121 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3181 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3241 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3301 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3361 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3421 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3481 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3541 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3601 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3661 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3721 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3781 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3841 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3901 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3961 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4021 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4081 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4141 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4201 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4261 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4321 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4381 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4441 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4501 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4561 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     4621 acgaggccct ttcgtcttca agaactgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     4681 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4741 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     4801 gtcacgtagc gatagcggag tgtactggct taactatgcg gcatcagagc agattgtact
     4861 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     4921 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4981 ttcgctatta cgccagcgcg gggaggcaga gattgcagta agctgagatc gcagcactgc
     5041 actccagcct gggcgacaga gtaagactct gtctcaaaaa taaaataaat aaatcaatca
     5101 gatattccaa tcttttcctt tatttattta tttattttct attttggaaa cacagtcctt
     5161 ccttattcca gaattacaca tatattctat ttttctttat atgctccagt tttttttaga
     5221 ccttcacctg aaatgtgtgt atacaaaatc taggccagtc cagcagagcc taaaggtaaa
     5281 aaataaaata ataaaaaata aataaaatct agctcactcc ttcacatcaa aatggagata
     5341 cagctgttag cattaaatac caaataaccc atcttgtcct caataatttt aagcgcctct
     5401 ctccaccaca tctaactcct gtcaaaggca tgtgcccctt ccgggcgctc tgctgtgctg
     5461 ccaaccaact ggcatgtgga ctctgcaggg tccctaactg ccaagcccca cagtgtgccc
     5521 tgaggctgcc ccttccttct agcggctgcc cccactcggc tttgctttcc ctagtttcag
     5581 ttacttgcgt tcagccaagg tctgaaacta ggtgcgcaca gagcggtaag actgcgagag
     5641 aaagagacca gctttacagg gggtttatca cagtgcaccc tgacagtcgt cagcctcaca
     5701 gggggtttat cacattgcac cctgacagtc gtcagcctca cagggggttt atcacagtgc
     5761 acccttacaa tcattccatt tgattcacaa tttttttagt ctctactgtg cctaacttgt
     5821 aagttaaatt tgatcagagg tgtgttccca gaggggaaaa cagtatatac agggttcagt
     5881 actatcgcat ttcagggtgg cccgagagct gcatccggag tatctagatg gagttccgcg
     5941 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     6001 cgtcaataat gactgatgtt cccatagtaa cgccaatagg gactttccat tgacgctaat
     6061 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc
     6121 ccgttgacgc aaatgggcgg taggcgtgta ctctagaagg tctatataag cagagctcgt
     6181 ttagtgaacc gtttatacta cttatctggt ctctctggtt agaccagatc tgagcctggg
     6241 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6301 tcaaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6361 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagt
     6421 aaagccagag gagatctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     6481 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     6541 agatgggtgc gagagcgtcg gtattaagcg ggggagaatt agataaatgg gaaaaaattc
     6601 ggttaaggcc agggggaaag aaacaatata aactaaaaca tatagtatgg gcaagcaggg
     6661 agctagaacg attcgcagtt aatcctggcc ttttagagac atcagaaggc tgtagacaaa
     6721 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     6781 atacaatagc agtcctctat tgtgtgcatc aaaggataga tgtaaaagac accaaggaag
     6841 ccttagataa gatagaggaa gagcaaaaca aaagtaagaa aaaggcacag caagcagcag
     6901 ctgacacagg aaacaacagc caggtcagcc aaaattaccc tatagtgcag aacctccagg
     6961 ggcaaatggt acatcaggcc atatcaccta gaactttaaa tgcatgggta aaagtagtag
     7021 aagagaaggc tttcagccca gaagtaatac ccatgttttc agcattatca gaaggagcca
     7081 ccccacaaga tttaaatacc atgctaaaca cagtgggggg acatcaagca gccatgcaaa
     7141 tgttaaaaga gaccatcaat gaggaagctg cagaatggga tagattgcat ccagtgcatg
     7201 cagggcctat tgcaccaggc cagatgagag aaccaagggg aagtgacata gcaggaacta
     7261 ctagcaaatt aagagaacaa tttggaaata ataaaacaat aatctttaag caatcctcag
     7321 gaggggaccc agaaattgta acgcacagtt ttaattgtgg aggggaattt ttctactgta
     7381 attcaacaca actgtttaat agtacttggt ttaatagtac ttggagtact gaagggtcaa
     7441 ataacactga aggaagtgac acaatcacac tcccatgcag aataaaacaa tttataaaca
     7501 tgtggcagga agtaggaaaa gcaatgtatg cccctcccat cagtggacaa attagatgtt
     7561 catcaaatat tactgggctg ctattaacaa gagatggtgg taataacaac aatgggtccg
     7621 agatcttcag acctggagga ggcgatatga gggacaattg gagaagtgaa ttatataaat
     7681 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     7741 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     7801 caggaagcac tatgggcgca cggtcaatga cgctgacggt acaggccaga caattattgt
     7861 ctgatatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     7921 tgcaactcac agtctggggc atcaaacagc tccaggcaag aatcctggct gtggaaagat
     7981 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     8041 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaataaca
     8101 tgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     8161 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     8221 gggcttgcgg ccggaattca caaatggcag tattcatcca caattttaaa agaaaagggg
     8281 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     8341 ctaaagaatt acaaaaacaa attactaaaa attcnaaaat tttcgggttt attacaggga
     8401 cagcagagat ccactttggg cggcctaggg ataacagggt aatgatatct ctgaccgcta
     8461 gcatcgtcga cccgggatcc gcggactagt gtacgcgtc
//
