LOCUS       sequence-329418-        7141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cre-inactivated hM3Dq-mCherry under control of the TRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7141)
  AUTHORS   Sabatini PV, Wang J, Rupp AC, Affinati AH, Flak JN, Li C, Olson DP, 
            Myers MG Jr
  TITLE     tTARGIT AAVs mediate the sensitive and flexible manipulation of 
            intersectional neuronal populations in mice.
  JOURNAL   eLife. 2021 Mar 11;10. pii: 66835. doi: 10.7554/eLife.66835.
  PUBMED    33704065
REFERENCE   2  (bases 1 to 7141)
  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: "eLife."; 
            date: "2021-03-11"; volume: "10. pii: 66835. doi"; pages: " 
            10.7554/eLife.66835"
COMMENT     Sequence Label: pAAV-TRE-DIO-hM3Dq-mCherry (Cre-OFF)
FEATURES             Location/Qualifiers
     source          1..7141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   531..671
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   542..671
                     /label=AAV2 ITR
     protein_bind    join(828..846,847..869,870..888,889..911,912..930,931..953,
                     954..972,973..995,996..1014,1015..1037,1038..1056,
                     1057..1079,1080..1098)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1131..1169
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    complement(1304..1337)
                     /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(1348..1381)
                     /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)."
     regulatory      3233..3242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3239..3949
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3384..3402)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3652..3671
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    3985..4018
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4029..4062
                     /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)."
     misc_feature    4086..4674
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4139..4159)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4557..4568)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4709..4726)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4715..4922
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4945..5074
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5136..5724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5216..5235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5895..6686,6687..6755))
                     /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     6518..6537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6756..6860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6984..7006)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7106..7125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca atagaccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga
      121 tagagttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctaag gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg
      481 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca
      541 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
      601 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      661 taggggttcc tgcggccgca cgcgaaacaa ttctgcagga attcggggcc gcggaggctg
      721 gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct ccaggcgatc
      781 tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc ctatcagtga
      841 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
      901 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
      961 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     1021 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     1081 ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga ggtaggcgtg
     1141 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
     1201 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc
     1261 ccgaattcga gctcggtacc cggggatcct ctagtcgacg tcgataactt cgtatagcat
     1321 acattatacg aagttatccg aatcgcaata acttcgtata ggatacttta tacgaagtta
     1381 tttggcgcgc caaatttaaa taacgcaacg taccttcgag ggggcccaag gaattcttac
     1441 ttgtacgatg accttgcaca ataacagtac aacctcgcct ttgtttccaa acatcagctc
     1501 ctcctggata cacagcccct ccgatgcagg gctgcccccg ggaaccgtca ctcatttcgg
     1561 cagctacaat gtttctcgag cagctggcaa tttctcctct ccagacggta ccaccgatga
     1621 ccctctggga ggtcataccg tctggcaagt ggtcttcatc gctttcttaa cgggcatcct
     1681 ggccttggtg accatcatcg gcaacatcct ggtaattgtg tcatttaagg tcaacaagca
     1741 gctgaagacg gtcaacaact acttcctctt aagcctggcc tgtgccgatc tgattatcgg
     1801 ggtcatttca atgaatctgt ttacgaccta catcatcatg aatcgatggg ccttagggaa
     1861 cttggcctgt gacctctggc ttgccattga ctgcgtagcc agcaatgcct ctgttatgaa
     1921 tcttctggtc atcagctttg acagatactt ttccatcacg aggccgctca cgtaccgagc
     1981 caaacgaaca acaaagagag ccggtgtgat gatcggtctg gcttgggtca tctcctttgt
     2041 cctttgggct cctgccatct tgttctggca atactttgtt ggaaagagaa ctgtgcctcc
     2101 gggagagtgc ttcattcagt tcctcagtga gcccaccatt acttttggca cagccatcgc
     2161 tggtttttat atgcctgtca ccattatgac tattttatac tggaggatct ataaggaaac
     2221 tgaaaagcgt accaaagagc ttgctggcct gcaagcctct gggacagagg cagagacaga
     2281 aaactttgtc caccccacgg gcagttctcg aagctgcagc agttacgaac ttcaacagca
     2341 aagcatgaaa cgctccaaca ggaggaagta tggccgctgc cacttctggt tcacaaccaa
     2401 gagctggaaa cccagctccg agcagatgga ccaagaccac agcagcagtg acagttggaa
     2461 caacaatgat gctgctgcct ccctggagaa ctccgcctcc tccgacgagg aggacattgg
     2521 ctccgagacg agagccatct actccatcgt gctcaagctt ccgggtcaca gcaccatcct
     2581 caactccacc aagttaccct catcggacaa cctgcaggtg cctgaggagg agctggggat
     2641 ggtggacttg gagaggaaag ccgacaagct gcaggcccag aagagcgtgg acgatggagg
     2701 cagttttcca aaaagcttct ccaagcttcc catccagcta gagtcagccg tggacacagc
     2761 taagacttct gacgtcaact cctcagtggg taagagcacg gccactctac ctctgtcctt
     2821 caaggaagcc actctggcca agaggtttgc tctgaagacc agaagtcaga tcactaagcg
     2881 gaaaaggatg tccctggtca aggagaagaa agcggcccag accctcagtg cgatcttgct
     2941 tgccttcatc atcacttgga ccccatacaa catcatggtt ctggtgaaca ccttttgtga
     3001 cagctgcata cccaaaacct tttggaatct gggctactgg ctgtgctaca tcaacagcac
     3061 cgtgaacccc gtgtgctatg ctctgtgcaa caaaacattc agaaccactt tcaagatgct
     3121 gctgctgtgc cagtgtgaca aaaaaaagag gcgcaagcag cagtaccagc agagacagtc
     3181 ggtcattttt cacaagcgcg cacccgagca ggccttgaag gatcccccgg tcgccaccat
     3241 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3301 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3361 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3421 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3481 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3541 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3601 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3661 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3721 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3781 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3841 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3901 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag ctagctagtt
     3961 ggcgcgccaa cctggccggc cttgataact tcgtataatg tatgctatac gaagttatcc
     4021 gaatcgcaat aacttcgtat aaagtatcct atacgaagtt atcgaagatc atcaagctta
     4081 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4141 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4201 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4261 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4321 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4381 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4441 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4501 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4561 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4621 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     4681 accgagcgct gctcgagaga tcgatctgcc tcgactgtgc cttctagttg ccagccatct
     4741 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4801 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4861 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4921 gacacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4981 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc
     5041 tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag
     5101 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5161 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5221 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5281 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5341 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5401 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5461 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5521 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5581 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5641 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5701 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5761 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5821 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5881 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5941 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6001 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6061 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6121 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6181 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6241 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6301 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6361 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6421 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6481 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6541 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6601 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6661 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6721 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6781 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6841 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6901 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6961 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7021 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7081 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     7141 a
//
