LOCUS       sequence-317851-        7462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of  hM3Dq-mCherry in a Flp-dependent fashion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7462)
  AUTHORS   Beier KT, Kim CK, Hoerbelt P, Hung LW, Heifets BD, DeLoach KE, Mosca
            TJ, Neuner S, Deisseroth K, Luo L, Malenka RC
  TITLE     Rabies screen reveals GPe control of cocaine-triggered plasticity.
  JOURNAL   Nature. 2017 Sep 21;549(7672):345-350. doi: 10.1038/nature23888. 
            Epub 2017 Sep 13.
  PUBMED    28902833
REFERENCE   2  (bases 1 to 7462)
  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: 
            "10.1038/nature23888"; journalName: "Nature"; date: "2017-09-21- 
            21"; volume: "549"; issue: "7672"; pages: "345-350"
COMMENT     Sequence Label: pAAV hSyn FLEx-FRT hM3Dq-mCherry
FEATURES             Location/Qualifiers
     source          1..7462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..48
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      1845..1854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1851..2561
                     /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(1996..2014)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2264..2283
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2677..2724)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(2802..3249)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3294..3423
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3485..4073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3565..3584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4244..5035,5036..5104))
                     /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     4867..4886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5105..5209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5333..5355)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5455..5474
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5491..5946)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5628..5649)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5840..5859
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6021..6161
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6032..6161
                     /label=AAV2 ITR
     polyA_signal    6201..6677
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6459..6478
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6709..7297
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6815..6826
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7224..7244
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcgg cgcgccgcca
       61 tgaccttgca caataacagt acaacctcgc ctttgtttcc aaacatcagc tcctcctgga
      121 tacacagccc ctccgatgca gggctgcccc cgggaaccgt cactcatttc ggcagctaca
      181 atgtttctcg agcagctggc aatttctcct ctccagacgg taccaccgat gaccctctgg
      241 gaggtcatac cgtctggcaa gtggtcttca tcgctttctt aacgggcatc ctggccttgg
      301 tgaccatcat cggcaacatc ctggtaattg tgtcatttaa ggtcaacaag cagctgaaga
      361 cggtcaacaa ctacttcctc ttaagcctgg cctgtgccga tctgattatc ggggtcattt
      421 caatgaatct gtttacgacc tacatcatca tgaatcgatg ggccttaggg aacttggcct
      481 gtgacctctg gcttgccatt gactgcgtag ccagcaatgc ctctgttatg aatcttctgg
      541 tcatcagctt tgacagatac ttttccatca cgaggccgct cacgtaccga gccaaacgaa
      601 caacaaagag agccggtgtg atgatcggtc tggcttgggt catctccttt gtcctttggg
      661 ctcctgccat cttgttctgg caatactttg ttggaaagag aactgtgcct ccgggagagt
      721 gcttcattca gttcctcagt gagcccacca ttacttttgg cacagccatc gctggttttt
      781 atatgcctgt caccattatg actattttat actggaggat ctataaggaa actgaaaagc
      841 gtaccaaaga gcttgctggc ctgcaagcct ctgggacaga ggcagagaca gaaaactttg
      901 tccaccccac gggcagttct cgaagctgca gcagttacga acttcaacag caaagcatga
      961 aacgctccaa caggaggaag tatggccgct gccacttctg gttcacaacc aagagctgga
     1021 aacccagctc cgagcagatg gaccaagacc acagcagcag tgacagttgg aacaacaatg
     1081 atgctgctgc ctccctggag aactccgcct cctccgacga ggaggacatt ggctccgaga
     1141 cgagagccat ctactccatc gtgctcaagc ttccgggtca cagcaccatc ctcaactcca
     1201 ccaagttacc ctcatcggac aacctgcagg tgcctgagga ggagctgggg atggtggact
     1261 tggagaggaa agccgacaag ctgcaggccc agaagagcgt ggacgatgga ggcagttttc
     1321 caaaaagctt ctccaagctt cccatccagc tagagtcagc cgtggacaca gctaagactt
     1381 ctgacgtcaa ctcctcagtg ggtaagagca cggccactct acctctgtcc ttcaaggaag
     1441 ccactctggc caagaggttt gctctgaaga ccagaagtca gatcactaag cggaaaagga
     1501 tgtccctggt caaggagaag aaagcggccc agaccctcag tgcgatcttg cttgccttca
     1561 tcatcacttg gaccccatac aacatcatgg ttctggtgaa caccttttgt gacagctgca
     1621 tacccaaaac cttttggaat ctgggctact ggctgtgcta catcaacagc accgtgaacc
     1681 ccgtgtgcta tgctctgtgc aacaaaacat tcagaaccac tttcaagatg ctgctgctgt
     1741 gccagtgtga caaaaaaaag aggcgcaagc agcagtacca gcagagacag tcggtcattt
     1801 ttcacaagcg cgcacccgag caggccttga aggatccccc ggtcgccacc atggtgagca
     1861 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     1921 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1981 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     2041 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     2101 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     2161 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     2221 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     2281 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     2341 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     2401 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     2461 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     2521 gccgccactc caccggcggc atggacgagc tgtacaagta agtcgacgaa gttcctatac
     2581 cttttgaaga ataggaactt cggaatagga acttctgtcg atggtgatgc ttggcaattc
     2641 gggtcgatgg tgaagcattg gtcttcctat tttgatgaag ttcctatact ttctagagaa
     2701 taggaacttc ggaataggaa cttcggtacc cccgggcccg cggtatcaaa tcaattcttt
     2761 gccaaaatga tgagacagca caggatccgg taccttctcg actgcgctct caggcacgac
     2821 acgactcctc cgctgcccac cgcagactga ggcagcgctg agtcgccggc gccgcagcgc
     2881 agatggtcgc gcccgtgccc ccctatctcg cgcctcgcgt ggtgcggtcc ggctgggccg
     2941 gcggcggcgc ggacgcgacc aaggtggccg ggaaggggag tttgcggggg accggcgagt
     3001 gacgtcagcg cgccttcagt gctgaggcgg cggtggcgcg cgccgccagg cgggggcgaa
     3061 ggcactgtcc gcggtgctga agctggcagt gcgcacgcgc ctcgccgcat cctgtttccc
     3121 ctccccctct ctgatagggg atgcgcaatt tggggaatgg gggttgggtg cttgtccagt
     3181 gggtcggggt cggtcgtcag gtaggcaccc ccaccccgcc tcatcctggt cctaaaaccc
     3241 acttgcactc atacgcaggg ccctctgcag tctagacaca cgcgtgcggc cgcaggaacc
     3301 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3361 accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3421 aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3481 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4381 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5281 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5341 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5401 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5461 cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat
     5521 tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa
     5581 tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca
     5641 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5701 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     5761 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5821 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa
     5881 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5941 gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6001 aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     6061 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     6121 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac
     6181 gtggttacct acaaaatcag aaggacaggg aagggagcag tggttcacgc ctgtaatccc
     6241 agcaatttgg gaggccaagg tgggtagatc acctgagatt aggagttgga gaccagcctg
     6301 gccaatatgg tgaaaccccg tctctaccaa aaaaacaaaa attagctgag cctggtcatg
     6361 catgcctgga atcccaacaa ctcgggaggc tgaggcagga gaatcgcttg aacccaggag
     6421 gcggagattg cagtgagcca agattgtgcc actgcactcc agcttggttc ccaatagacc
     6481 ccgcaggccc tacaggttgt cttcccaact tgccccttgc tccataccac ccccctccac
     6541 cccataatat tatagaagga cacctagtca gacaaaatga tgcaacttaa ttttattagg
     6601 acaaggctgg tgggcactgg agtggcaact tccagggcca ggagaggcac tggggagggg
     6661 tcacagggat gccacccgta gatctctcga gcagcgctcg gtatcgatgc ggggaggcgg
     6721 cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga ggccgcagag
     6781 ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg acgtagcaga
     6841 aggacgtccc gcgcagaatc caggtggcaa cataggcgag cagccaagga aaggacgatg
     6901 atttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc
     6961 gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa gcgaaagtcc
     7021 cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca
     7081 cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa agagacagca
     7141 accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca
     7201 aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag
     7261 tcaatctttc acaaattttg taatccagag gttgattatc gataagcttg atatcgaatt
     7321 catcgatcgg ccgcatataa gcttgaagtt cctattccga agttcctatt cttcaaaagg
     7381 tataggaact tcgggtcgat ggtgagatct ggactagagg gtcgatggtg atgcttggat
     7441 caaaatagga agaatcccaa tg
//
