LOCUS       sequence-308812-        6982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent hM4D(Gi) DREADD-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6982)
  AUTHORS   Runegaard AH, Sorensen AT, Fitzpatrick CM, Jorgensen SH, Petersen 
            AV, Hansen NW, Weikop P, Andreasen JT, Mikkelsen JD, Perrier JF, 
            Woldbye D, Rickhag M, Wortwein G, Gether U
  TITLE     Locomotor- and Reward-Enhancing Effects of Cocaine Are 
            Differentially Regulated by Chemogenetic Stimulation of Gi-Signaling
            in Dopaminergic Neurons.
  JOURNAL   eNeuro. 2018 Jun 18;5(3). pii: eN-NWR-0345-17. doi: 
            10.1523/ENEURO.0345-17.2018. eCollection 2018 May-Jun.
  PUBMED    29938215
REFERENCE   2  (bases 1 to 6982)
  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: "eNeuro."; 
            date: "2018-06-18"; volume: "5(3). pii: eN-NWR-0345-17. doi"; pages:
            " 10.1523/ENEURO.0345-17.2018. eCollection 2018 May-Jun"
COMMENT     Sequence Label: pAAV-hSyn-fDIO-hM4D(Gi)-mCherry-WPREpA
FEATURES             Location/Qualifiers
     source          1..6982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        137..584
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    622..655
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(748..1458)
                     /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(1026..1045)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1295..1313
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1455..1464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2913..2922
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2929..2962)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3071..3659
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3124..3144)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3542..3553)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3691..4167
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3890..3909)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4207..4347
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4207..4336
                     /label=AAV2 ITR
     rep_origin      4422..4877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4509..4528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4719..4740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4894..4913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5013..5035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5073..5091)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5159..5263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5264..5332,5333..6124)
                     /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(5482..5501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6295..6883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6784..6803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6945..6982,1..92)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
       61 gggagtggcc aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca
      121 gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg
      181 cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg
      241 cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc
      301 cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc
      361 tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc
      421 accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga
      481 taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt
      541 ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga
      601 tcctctagag tcgactccgg agaagttcct attctctaga aagtatagga acttcgcaga
      661 atggtagctg gattgtagct gctattagca atatgaaacc tcttagaagt tcctattctt
      721 caaaaggtat aggaacttcg gcgcgcctta cttgtacagc tcgtccatgc cgccggtgga
      781 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
      841 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
      901 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
      961 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     1021 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     1081 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     1141 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     1201 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     1261 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
     1321 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     1381 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
     1441 ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgc ctggcagtgc cgatgttccg
     1501 atactggcac agcagcaggt gccggaaggt ctttttaaag gtggcgttgc acagagcata
     1561 gcaggcaggg ttgatggtgc tgttgacgta gcagagccag tagccaatgg accacaccgt
     1621 gtcagggatg cagctctggc agaaggtgtt caccaggacc atgacgttgt agggcgtcca
     1681 ggtgaggatg aaggccagca gaatggcaaa gatcgttcgt gtcactttgc gctcccgggc
     1741 cgccatctgc cgcttcttgc gcacctggtt gcgagcgatg ctggcgaact tgcgggccac
     1801 gttggccgca gggcgcatgc cagccggcgt ggcaggcaca atctcaatgg ctgtcacaca
     1861 ctcattgcct gtctgcttcg tcacaatctg gatcttggac catctggagg ctgggttgag
     1921 ggcccgcggc tgcaggggag gggcgggcat ggcgggcgtg gtggcctctg tggtggacag
     1981 ctctgtggct gggcgttcct tggtgttctg ggtggcactg cctgagctgg actcattgga
     2041 agtgtcctta tcagccacgg ggcgcggtgg cggtggcagc gctggcgggg gggcctcctc
     2101 cagcttgcca ttgcgcagct cctcccgggc ggcctccccg ggcgggggct tcttgacgct
     2161 ctgcttcatt agtgggctct tgaggaaggc cagcgtcttg gctttcttct ccttcgggcc
     2221 ctcgggccgg tgcttgtgga ctcggctgcg actggccagg gagatgtgga tgtacagcac
     2281 cgtcatgatg accacaggca ggtagaagcc agcaatggct gtgccaaagg tcactgctgg
     2341 gttggacagg aactggatga agcactggtt gtcgggcacc gtccgcttac ccaccacaaa
     2401 ctgccagaac aagatggcag gcgcccagag cacgaaggac agtacccagg cagcagcaat
     2461 catgaggcct gccatcttgg tggtgcgccg ggcagggtag gtgagaggct tggtgacgca
     2521 gaagtagcgg tcaaagctga tgatgagaag gttcatgacg gaggcgttgc tcaccacgca
     2581 gtccagggcc agccacaggt cgcagaccac ggcgcccagg ggccagtagc ccttgatgat
     2641 gtacacggtg tagaggttca tggagaaggc gcctatgatg agatcagcac acgccaggct
     2701 gaagaggaag tagttgttga ctgtctgcag ctgcctgttg accttgatgg acagcatcac
     2761 caggatgttg cccacgacag tcaccaggct cagggagcct gtcactgtgg caatgaagac
     2821 catttccacc gtctcatagc gattgtggga tgatgacgtg accaggcgca cggactgatt
     2881 gcccgagctg ccattgacag gtgtgaagtt ggccatggtg gcgctagcga agttcctata
     2941 ctttctagag aataggaact tcttgcctta acccagaaat tatcactgtt attctttaga
     3001 atggtgcaaa gagaagttcc tatacctttt gaagaatagg aacttcgaat tcgatatcaa
     3061 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3121 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3181 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3241 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3301 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3361 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3421 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3481 ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3541 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3601 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3661 tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg
     3721 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     3781 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     3841 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     3901 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     3961 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     4021 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     4081 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     4141 tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc
     4201 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4261 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4321 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4381 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4621 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4741 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4801 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4861 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4921 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4981 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5041 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5101 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5941 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6421 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     6961 cgctcgctcg ctcactgagg cc
//
