LOCUS       sequence-343349-        7191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-off TVA-mCherry fusion protein with 66T mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7191)
  AUTHORS   Melzer S, Newmark ER, Mizuno GO, Hyun M, Philson AC, Quiroli E, 
            Righetti B, Gregory MR, Huang KW, Levasseur J, Tian L, Sabatini BL
  TITLE     Bombesin-like peptide recruits disinhibitory cortical circuits and 
            enhances fear memories.
  JOURNAL   Cell. 2021 Oct 28;184(22):5622-5634.e25. doi: 
            10.1016/j.cell.2021.09.013. Epub 2021 Oct 4.
  PUBMED    34610277
REFERENCE   2  (bases 1 to 7191)
  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"; doi: 
            "10.1016/j.cell.2021.09.013"; journalName: "Cell"; date: 
            "2021-10-28- 28"; volume: "184"; issue: "22"; pages: "5622-5634.e25"
COMMENT     Sequence Label: pAAV CAG-DO-TC66T-mCherry
FEATURES             Location/Qualifiers
     source          1..7191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          128..1144
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1068..1090
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1152..1171
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1224..1257
                     /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    1320..1353
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1801..2508
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1943..1961)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2211..2230
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2518..2551)
                     /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    complement(2622..2655)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2679..3267
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2732..2752)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3150..3161)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3291..3767
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3490..3509)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3813..3953
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3813..3942
                     /label=AAV2 ITR
     rep_origin      4028..4483
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4115..4134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4325..4346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4500..4519)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4619..4641
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4679..4697)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4765..4869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4870..4938,4939..5730)
                     /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     complement(5088..5107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5901..6489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6390..6409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6551..6680
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        6770..7073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
       61 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      121 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      181 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      241 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      301 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
      361 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
      421 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
      481 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
      541 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
      601 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
      661 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
      721 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
      781 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
      841 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
      901 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
      961 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1021 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1081 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
     1141 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgat
     1201 ttgataccgc gggcccggcc gggataactt cgtataaagt atcctatacg aagttatatc
     1261 aaaataggaa gaccaatgct tcaccatcga cccgaattgc caagcatcac catcgacaca
     1321 taacttcgta taatgtatgc tatacgaagt tatgtcgaca ccatggcgcg gctgctgccc
     1381 gcgctgctgc tgctgctgct gcccggtaac gtgaccggta acgggtccgg taacggttct
     1441 ttgtcccgtt gcccccccgg tcagttccgc tgctcggagc cgcccggtgc ccacggggag
     1501 tgttacccgc aggactggct gtgcgacgga caccccgact gcgacgacgg gcgggacacc
     1561 tggggctgcg ggaccagcgc gatccccgcg gtgcccacgg acaacggcac agaggctccc
     1621 actgcccctg ctcctggacg tgctctgcca gccaggaatc acggccgcat gtggatgctg
     1681 atcactgcag tgctcctgtg ctgcctggta gctgtgggtg gtatcgctgc atgggggaag
     1741 tccaaagcaa aaagcaggtc tgacatcttc agtcttgaaa gcgcatccaa ggggggtggg
     1801 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     1861 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1921 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     1981 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2041 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2101 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2161 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2221 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2281 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2341 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2401 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     2461 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagg cgcgcctata
     2521 acttcgtata ggatacttta tacgaagtta tcattgggat tcttcctatt ttgatccaag
     2581 catcaccatc gaccctctag tccagatctc accatcgacc cataacttcg tatagcatac
     2641 attatacgaa gttataagct tatatgcggc cgatcgataa tcaacctctg gattacaaaa
     2701 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2761 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2821 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2881 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2941 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3001 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3061 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     3121 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3181 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3241 ggatctccct ttgggccgcc tccccgcatc gataccgtag atcgatctac gggtggcatc
     3301 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     3361 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     3421 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     3481 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     3541 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     3601 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     3661 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     3721 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
     3781 taaccacgtg cggaccgaac tagtgcggcc gcaggaaccc ctagtgatgg agttggccac
     3841 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3901 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3961 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     4021 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4081 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4141 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4201 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4261 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4321 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4381 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4441 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4501 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4561 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4621 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4681 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4741 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4801 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4861 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4921 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4981 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5041 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5101 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5161 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5221 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5281 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5341 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5401 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5461 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5521 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5581 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5641 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5701 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5761 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5821 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5881 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5941 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6001 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6061 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6121 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6181 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6241 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6301 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6361 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6421 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6481 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6541 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     6601 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     6661 ctccatcact aggggttcct gcggccgcac gcgtaattcc cggggaatct agttattaat
     6721 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     6781 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     6841 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6901 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6961 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     7021 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     7081 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     7141 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg g
//
