LOCUS       sequence-329449-        7130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cre-dependent hM3Dq-mCherry under control of the TRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7130)
  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 7130)
  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
FEATURES             Location/Qualifiers
     source          1..7130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(42..60,61..83,84..102,103..125,126..144,145..167,
                     168..186,187..209,210..228,229..251,252..270,271..293,
                     294..312)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        345..383
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    complement(518..551)
                     /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(562..595)
                     /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)."
     CDS             complement(651..1361)
                     /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(929..948)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1198..1216
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1358..1367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3188..3221
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3232..3265
                     /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    3289..3877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3342..3362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3760..3771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3912..3929)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3918..4125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4148..4277
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4339..4927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4419..4438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5098..5889,5890..5958))
                     /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     5721..5740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5959..6063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6187..6209)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6309..6328
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6345..6800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6482..6503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6694..6713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6875..7015
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6886..7015
                     /label=AAV2 ITR
ORIGIN
        1 ttcactaaac gagctctgct tatataggtc gagtttacca ctccctatca gtgatagaga
       61 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
      121 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
      181 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
      241 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
      301 cagtgataga gaaaagtgaa agtcgagctc ggtacccggg tcgaggtagg cgtgtacggt
      361 gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      421 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc ggccccgaat
      481 tcgagctcgg tacccgggga tcctctagtc gacgtcgata acttcgtata gcatacatta
      541 tacgaagtta tccgaatcgc aataacttcg tataggatac tttatacgaa gttatttggc
      601 gcgccaaatt taaataacgc aacgtacctt cgagggggcc caaggaattc ttacttgtac
      661 agctcgtcca tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg
      721 gtgtagtcct cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc
      781 agctgcacgg gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg
      841 tccttcagct tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc
      901 cgctcggagg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg
      961 aagttggtgc cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc
     1021 tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc
     1081 tcggggaagg acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg
     1141 gagccgtaca tgaactgagg ggacaggatg tcccaggcga agggcagggg gccacccttg
     1201 gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg
     1261 atctcgaact cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc
     1321 ttgatgatgg ccatgttatc ctcctcgccc ttgctcacca tggtggcgac cgggggatcc
     1381 ttcaaggcct gctcgggtgc gcgcttgtga aaaatgaccg actgtctctg ctggtactgc
     1441 tgcttgcgcc tctttttttt gtcacactgg cacagcagca gcatcttgaa agtggttctg
     1501 aatgttttgt tgcacagagc atagcacacg gggttcacgg tgctgttgat gtagcacagc
     1561 cagtagccca gattccaaaa ggttttgggt atgcagctgt cacaaaaggt gttcaccaga
     1621 accatgatgt tgtatggggt ccaagtgatg atgaaggcaa gcaagatcgc actgagggtc
     1681 tgggccgctt tcttctcctt gaccagggac atccttttcc gcttagtgat ctgacttctg
     1741 gtcttcagag caaacctctt ggccagagtg gcttccttga aggacagagg tagagtggcc
     1801 gtgctcttac ccactgagga gttgacgtca gaagtcttag ctgtgtccac ggctgactct
     1861 agctggatgg gaagcttgga gaagcttttt ggaaaactgc ctccatcgtc cacgctcttc
     1921 tgggcctgca gcttgtcggc tttcctctcc aagtccacca tccccagctc ctcctcaggc
     1981 acctgcaggt tgtccgatga gggtaacttg gtggagttga ggatggtgct gtgacccgga
     2041 agcttgagca cgatggagta gatggctctc gtctcggagc caatgtcctc ctcgtcggag
     2101 gaggcggagt tctccaggga ggcagcagca tcattgttgt tccaactgtc actgctgctg
     2161 tggtcttggt ccatctgctc ggagctgggt ttccagctct tggttgtgaa ccagaagtgg
     2221 cagcggccat acttcctcct gttggagcgt ttcatgcttt gctgttgaag ttcgtaactg
     2281 ctgcagcttc gagaactgcc cgtggggtgg acaaagtttt ctgtctctgc ctctgtccca
     2341 gaggcttgca ggccagcaag ctctttggta cgcttttcag tttccttata gatcctccag
     2401 tataaaatag tcataatggt gacaggcata taaaaaccag cgatggctgt gccaaaagta
     2461 atggtgggct cactgaggaa ctgaatgaag cactctcccg gaggcacagt tctctttcca
     2521 acaaagtatt gccagaacaa gatggcagga gcccaaagga caaaggagat gacccaagcc
     2581 agaccgatca tcacaccggc tctctttgtt gttcgtttgg ctcggtacgt gagcggcctc
     2641 gtgatggaaa agtatctgtc aaagctgatg accagaagat tcataacaga ggcattgctg
     2701 gctacgcagt caatggcaag ccagaggtca caggccaagt tccctaaggc ccatcgattc
     2761 atgatgatgt aggtcgtaaa cagattcatt gaaatgaccc cgataatcag atcggcacag
     2821 gccaggctta agaggaagta gttgttgacc gtcttcagct gcttgttgac cttaaatgac
     2881 acaattacca ggatgttgcc gatgatggtc accaaggcca ggatgcccgt taagaaagcg
     2941 atgaagacca cttgccagac ggtatgacct cccagagggt catcggtggt accgtctgga
     3001 gaggagaaat tgccagctgc tcgagaaaca ttgtagctgc cgaaatgagt gacggttccc
     3061 gggggcagcc ctgcatcgga ggggctgtgt atccaggagg agctgatgtt tggaaacaaa
     3121 ggcgaggttg tactgttatt gtgcaaggtc atgctagcta gttggcgcgc caacctggcc
     3181 ggccttgata acttcgtata atgtatgcta tacgaagtta tccgaatcgc aataacttcg
     3241 tataaagtat cctatacgaa gttatcgaag atcatcaagc ttatcgataa tcaacctctg
     3301 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3361 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3421 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     3481 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     3541 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     3601 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     3661 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     3721 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3781 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3841 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     3901 agatcgatct gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3961 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4021 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4081 cagcaagggg gaggattggg aagacaatag caggcatgct ggggacacgt gcggaccgag
     4141 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4201 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc
     4261 gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4741 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4921 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5041 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5101 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5161 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5221 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5281 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5341 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5401 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5461 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5521 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5581 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5641 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5701 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5761 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5821 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5881 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5941 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6001 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6061 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     6121 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     6181 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6241 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     6301 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
     6361 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaatagac
     6421 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt tgagtgttgt
     6481 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     6541 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     6601 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     6661 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6721 taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6781 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     6841 agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg cgctcgctcg
     6901 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     6961 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     7021 cgcacgcgaa acaattctgc aggaattcgg ggccgcggag gctggatcgg tcccggtgtc
     7081 ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg
//
