LOCUS       sequence-374532-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent TRE-tTA expression under the control of Syn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  TITLE     AAV expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5520)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-Syn-Flex-TREx2-tTA
FEATURES             Location/Qualifiers
     source          1..5520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      911..1499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1400..1419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   1736..1865
                     /label=AAV2 ITR
     polyA_signal    2027..2148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2064..2083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2118..2137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    2191..2779
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2297..2308
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2706..2726
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2821..2854
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2925..2958
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3016..3034
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3052..3070
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3780..3890
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     protein_bind    complement(3925..3958)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(4021..4054)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(4084..4531)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4613..4742
                     /label=AAV2 ITR
     rep_origin      4814..5269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4901..4920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5111..5132
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5295..5399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5400..5468,5469..5520,1..740)
                     /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"
ORIGIN
        1 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
       61 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      121 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      181 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      241 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      301 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      361 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      421 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      481 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      541 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      601 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      661 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      721 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      781 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      841 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      901 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      961 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1021 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1381 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1441 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1501 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1681 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
     1741 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     1801 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     1861 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
     1921 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1981 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
     2161 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
     2221 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
     2281 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
     2341 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
     2401 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     2461 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
     2521 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
     2581 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
     2641 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     2701 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     2761 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcgaacctc ttcgagggac
     2821 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     2881 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     2941 tatcctatac gaagttatac taggatccgc caccatgact agtggatcct ctaaaacagg
     3001 taagtcccat taatctccct atcagtgata gagaaggtct gaagagttta ctccctatca
     3061 gtgatagaga ttaattcctc tactaacctt gttcatcttt tctttttttt tctacaggtc
     3121 ctgggtgatt aacagcttaa gggcccgcca ccatgagtcg gctggataaa tctaaagtca
     3181 taaacggcgc tctggaatta ctcaatgaag tcggtatcga aggcctgacg acaaggaaac
     3241 tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg
     3301 ccctgctcga tgccctggcc atcgagatgc tggacaggca tcatacccac ttctgccccc
     3361 tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc
     3421 tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt
     3481 acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg
     3541 cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg
     3601 agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc
     3661 tgagacaagc aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc
     3721 tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg
     3781 acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc
     3841 ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaag
     3901 aattcaagct ttgaaagctc tagtataact tcgtatagca tacattatac gaagttatgg
     3961 gtcgatggtg atgcttggca attcgggtcg atggtgaagc attggtcttc ctattttgat
     4021 ataacttcgt ataggatact ttatacgaag ttatggggat ccttgctagc agcttgaatt
     4081 cgactgcgct ctcaggcacg acacgactcc tccgctgccc accgcagact gaggcagcgc
     4141 tgagtcgccg gcgccgcagc gcagatggtc gcgcccgtgc ccccctatct cgcgcctcgc
     4201 gtggtgcggt ccggctgggc cggcggcggc gcggacgcga ccaaggtggc cgggaagggg
     4261 agtttgcggg ggaccggcga gtgacgtcag cgcgccttca gtgctgaggc ggcggtggcg
     4321 cgcgccgcca ggcgggggcg aaggcactgt ccgcggtgct gaagctggca gtgcgcacgc
     4381 gcctcgccgc atcctgtttc ccctccccct ctctgatagg ggatgcgcaa tttggggaat
     4441 gggggttggg tgcttgtcca gtgggtcggg gtcggtcgtc aggtaggcac ccccaccccg
     4501 cctcatcctg gtcctaaaac ccacttgcac tcatacgcag ggccctctgc agagatcttc
     4561 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     4621 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4681 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4741 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4801 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4861 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4921 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4981 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5041 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5101 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5161 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5221 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     5281 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5341 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5401 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5461 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
//
