LOCUS       sequence-181128-        7345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gi-coupled hM4D-IRES-mCitrine under the control of human synapsin 
            promote.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7345)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7345)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn-HA-hM4D(Gi)-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..7345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             663..689
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2145..2718
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2312..2329)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2539..2558
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2719..2728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2725..2727,2728..2730,2731..3441)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2770..2791)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3031..3050)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3378..3399
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3472..4060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3525..3545)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3943..3954)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4092..4568
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4291..4310)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4608..4748
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4608..4737
                     /label=AAV2 ITR
     rep_origin      4823..5278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4910..4929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5120..5141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5295..5314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5414..5436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5474..5492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5560..5664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5665..5733,5734..6525)
                     /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(5883..5902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6696..7284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7185..7204
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gacgccacca
      661 tgtacccata cgatgttcca gattacgcta tggccaactt cacacctgtc aatggcagct
      721 cgggcaatca gtccgtgcgc ctggtcacgt catcatccca caatcgctat gagacggtgg
      781 aaatggtctt cattgccaca gtgacaggct ccctgagcct ggtgactgtc gtgggcaaca
      841 tcctggtgat gctgtccatc aaggtcaaca ggcagctgca gacagtcaac aactacttcc
      901 tcttcagcct ggcgtgtgct gatctcatca taggcgcctt ctccatgaac ctctacaccg
      961 tgtacatcat caagggctac tggcccctgg gcgccgtggt ctgcgacctg tggctggccc
     1021 tggactgcgt ggtgagcaac gcctccgtca tgaaccttct catcatcagc tttgaccgct
     1081 acttctgcgt caccaagcct ctcacctacc ctgcccggcg caccaccaag atggcaggcc
     1141 tcatgattgc tgctgcctgg gtactgtcct tcgtgctctg ggcgcctgcc atcttgttct
     1201 ggcagtttgt ggtgggtaag cggacggtgc ccgacaacca gtgcttcatc cagttcctgt
     1261 ccaacccagc agtgaccttt ggcacagcca ttgctggctt ctacctgcct gtggtcatca
     1321 tgacggtgct gtacatccac atctccctgg ccagtcgcag ccgagtccac aagcaccggc
     1381 ccgagggccc gaaggagaag aaagccaaga cgctggcctt cctcaagagc ccactaatga
     1441 agcagagcgt caagaagccc ccgcccgggg aggccgcccg ggaggagctg cgcaatggca
     1501 agctggagga ggcccccccg ccagcgctgc caccgccacc gcgccccgtg gctgataagg
     1561 acacttccaa tgagtccagc tcaggcagtg ccacccagaa caccaaggaa cgcccagcca
     1621 cagagctgtc caccacagag gccaccacgc ccgccatgcc cgcccctccc ctgcagccgc
     1681 gggccctcaa cccagcctcc agatggtcca agatccagat tgtgacgaag cagacaggca
     1741 atgagtgtgt gacagccatt gagattgtgc ctgccacgcc ggctggcatg cgccctgcgg
     1801 ccaacgtggc ccgcaagttc gccagcatcg ctcgcaacca ggtgcgcaag aagcggcaga
     1861 tggcggcccg ggagcgcaaa gtgacacgaa cgatctttgc cattctgctt gccttcatcc
     1921 tcacctggac gccctacaac gtcatggtcc tggtgaacac cttctgccag agctgcatcc
     1981 ctgacacggt gtggtccatt ggctactggc tctgctacgt caacagcacc atcaaccctg
     2041 cctgctatgc tctgtgcaac gccaccttta aaaagacctt ccggcacctg ctgctgtgcc
     2101 agtatcggaa catcggcact gccaggtagg cggccgcgat atcgcccctc tccctccccc
     2161 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     2221 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     2281 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     2341 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     2401 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     2461 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     2521 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     2581 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     2641 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     2701 tgaaaaacac gatgataagc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2761 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2821 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2881 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gatgtgcttc
     2941 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3001 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3061 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3121 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3181 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3241 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3301 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccaaactgag caaagacccc
     3361 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3421 ggcatggacg agctgtacaa gtgataggaa ttcgatatca agcttatcga taatcaacct
     3481 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3541 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3601 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3661 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3721 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3781 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3841 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3901 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3961 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     4021 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     4081 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     4141 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     4201 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     4261 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     4321 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     4381 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     4441 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     4501 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     4561 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4621 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4681 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4741 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4801 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4861 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4921 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4981 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     5041 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5101 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     5161 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     5221 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     5281 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5341 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5401 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5461 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5521 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5581 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5641 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5701 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5761 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5821 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5881 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5941 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6001 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6061 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6121 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6181 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6241 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6301 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6361 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6421 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6481 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6541 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6841 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7321 ttttgctggc cttttgctca catgt
//
