LOCUS       sequence-442270-        7018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gq coupled DREADD hM3D under the rat 
            pro-melanin-concentrating hormone promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   Wang Y, Guo R, Chen B, Rahman T, Cai L, Li Y, Dong Y, Tseng GC, Fang
            J, Seney ML, Huang YH
  TITLE     Cocaine-induced neural adaptations in the lateral hypothalamic 
            melanin-concentrating hormone neurons and the role in regulating 
            rapid eye movement sleep after withdrawal.
  JOURNAL   Mol Psychiatry. 2021 Jul;26(7):3152-3168. doi: 
            10.1038/s41380-020-00921-1. Epub 2020 Oct 22.
  PUBMED    33093653
REFERENCE   2  (bases 1 to 7018)
  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/s41380-020-00921-1"; journalName: "Mol Psychiatry"; date: 
            "2021-07"; volume: "26"; issue: "7"; pages: "3152-3168"
COMMENT     Sequence Label: pAAV-MCHpr-DREADD (Gq)-mCherry
FEATURES             Location/Qualifiers
     source          1..7018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2404..2413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2410..3120
                     /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(2555..2573)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2823..2842
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3145..3733
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3198..3218)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3616..3627)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3765..4241
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3964..3983)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4281..4421
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4281..4410
                     /label=AAV2 ITR
     rep_origin      4496..4951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4583..4602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4793..4814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4968..4987)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5087..5109
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5233..5337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5338..5406,5407..6198)
                     /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(5556..5575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6369..6957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6858..6877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttctaga gataacttct atttaataag gattaaaagg
      181 gcataaagct tttatcctta ttatccatga attaaataac acgtcttaat atcttaatcg
      241 gaaagtacat agaaggtaac tgtaatattg ggagagcctg gtgtcctatg gtcttcctta
      301 ataacatccc actttcaaga ggctataatc tactcacaga aagccttctc tcaaagacct
      361 taatcagatt ctgggaaaat tcacacacga aataaacaac aggccactaa atccaaaaca
      421 atgcaagcat tagcagcttt acaaaggcga ggctctacgg ggtgattcat ttctaaaaag
      481 aaaagataag gccttcaagt gttttctatt caggcacaaa gtatataaag gtaggaatca
      541 ttcagtcgcc agcagagtgt ggcattctcc ccacattctc ttcggcttta cggagcagca
      601 aacagggtcg acgccaccat gaccttgcac aataacagta caacctcgcc tttgtttcca
      661 aacatcagct cctcctggat acacagcccc tccgatgcag ggctgccccc gggaaccgtc
      721 actcatttcg gcagctacaa tgtttctcga gcagctggca atttctcctc tccagacggt
      781 accaccgatg accctctggg aggtcatacc gtctggcaag tggtcttcat cgctttctta
      841 acgggcatcc tggccttggt gaccatcatc ggcaacatcc tggtaattgt gtcatttaag
      901 gtcaacaagc agctgaagac ggtcaacaac tacttcctct taagcctggc ctgtgccgat
      961 ctgattatcg gggtcatttc aatgaatctg tttacgacct acatcatcat gaatcgatgg
     1021 gccttaggga acttggcctg tgacctctgg cttgccattg actgcgtagc cagcaatgcc
     1081 tctgttatga atcttctggt catcagcttt gacagatact tttccatcac gaggccgctc
     1141 acgtaccgag ccaaacgaac aacaaagaga gccggtgtga tgatcggtct ggcttgggtc
     1201 atctcctttg tcctttgggc tcctgccatc ttgttctggc aatactttgt tggaaagaga
     1261 actgtgcctc cgggagagtg cttcattcag ttcctcagtg agcccaccat tacttttggc
     1321 acagccatcg ctggttttta tatgcctgtc accattatga ctattttata ctggaggatc
     1381 tataaggaaa ctgaaaagcg taccaaagag cttgctggcc tgcaagcctc tgggacagag
     1441 gcagagacag aaaactttgt ccaccccacg ggcagttctc gaagctgcag cagttacgaa
     1501 cttcaacagc aaagcatgaa acgctccaac aggaggaagt atggccgctg ccacttctgg
     1561 ttcacaacca agagctggaa acccagctcc gagcagatgg accaagacca cagcagcagt
     1621 gacagttgga acaacaatga tgctgctgcc tccctggaga actccgcctc ctccgacgag
     1681 gaggacattg gctccgagac gagagccatc tactccatcg tgctcaagct tccgggtcac
     1741 agcaccatcc tcaactccac caagttaccc tcatcggaca acctgcaggt gcctgaggag
     1801 gagctgggga tggtggactt ggagaggaaa gccgacaagc tgcaggccca gaagagcgtg
     1861 gacgatggag gcagttttcc aaaaagcttc tccaagcttc ccatccagct agagtcagcc
     1921 gtggacacag ctaagacttc tgacgtcaac tcctcagtgg gtaagagcac ggccactcta
     1981 cctctgtcct tcaaggaagc cactctggcc aagaggtttg ctctgaagac cagaagtcag
     2041 atcactaagc ggaaaaggat gtccctggtc aaggagaaga aagcggccca gaccctcagt
     2101 gcgatcttgc ttgccttcat catcacttgg accccataca acatcatggt tctggtgaac
     2161 accttttgtg acagctgcat acccaaaacc ttttggaatc tgggctactg gctgtgctac
     2221 atcaacagca ccgtgaaccc cgtgtgctat gctctgtgca acaaaacatt cagaaccact
     2281 ttcaagatgc tgctgctgtg ccagtgtgac aaaaaaaaga ggcgcaagca gcagtaccag
     2341 cagagacagt cggtcatttt tcacaagcgc gcacccgagc aggccttgaa ggatcccccg
     2401 gtcgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     2461 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     2521 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     2581 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     2641 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     2701 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     2761 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     2821 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     2881 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     2941 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     3001 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     3061 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     3121 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3181 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3241 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3301 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3361 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3421 ggaactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3481 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3541 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     3601 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3661 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3721 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     3781 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     3841 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3901 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3961 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     4021 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     4081 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     4141 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     4201 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     4261 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4321 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4381 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4441 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4501 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4561 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4621 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4681 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4741 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4801 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4861 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4921 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4981 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5041 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5101 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5161 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5221 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5581 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5881 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6481 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
