LOCUS       sequence-75489-e        7888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gq-coupled hM3D-IRES-mCitrine under the control of GFAP promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7888)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7888)
  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            "Unpublished"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-GFAP-HA-hM3D(Gq)-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..7888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     CDS             876..902
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2688..3261
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2855..2872)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3082..3101
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3262..3271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3268..3270,3271..3273,3274..3984)
                     /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(3313..3334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3574..3593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3921..3942
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4015..4603
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4068..4088)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4486..4497)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4635..5111
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4834..4853)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5151..5291
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5151..5280
                     /label=AAV2 ITR
     rep_origin      5366..5821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5453..5472)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5663..5684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5838..5857)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5957..5979
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6103..6207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6208..6276,6277..7068)
                     /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"
     rep_origin      7239..7827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7728..7747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagagatcta acatatcctg
      181 gtgtggagta ggggacgctg ctctgacaga ggctcggggg cctgagctgg ctctgtgagc
      241 tggggaggag gcagacagcc aggccttgtc tgcaagcaga cctggcagca ttgggctggc
      301 cgccccccag ggcctcctct tcatgcccag tgaatgactc accttggcac agacacaatg
      361 ttcggggtgg gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa atgccttccg
      421 agaagcccat tgagcagggg gcttgcattg caccccagcc tgacagcctg gcatcttggg
      481 ataaaagcag cacagccccc taggggctgc ccttgctgtg tggcgccacc ggcggtggag
      541 aacaaggctc tattcagcct gtgcccagga aaggggatca ggggatgccc aggcatggac
      601 agtgggtggc agggggggag aggagggctg tctgcttccc agaagtccaa ggacacaaat
      661 gggtgagggg agagctctcc ccatagctgg gctgcggccc aaccccaccc cctcaggcta
      721 tgccaggggg tgttgccagg ggcacccggg catcgccagt ctagcccact ccttcataaa
      781 gccctcgcat cccaggagcg agcagagcca gagcaggttg gagaggagac gcatcacctc
      841 cgctgctcgc ggggtctaga gtcgacgcca ccatgtaccc atacgatgtt ccagattacg
      901 ctaccttgca caataacagt acaacctcgc ctttgtttcc aaacatcagc tcctcctgga
      961 tacacagccc ctccgatgca gggctgcccc cgggaaccgt cactcatttc ggcagctaca
     1021 atgtttctcg agcagctggc aatttctcct ctccagacgg taccaccgat gaccctctgg
     1081 gaggtcatac cgtctggcaa gtggtcttca tcgctttctt aacgggcatc ctggccttgg
     1141 tgaccatcat cggcaacatc ctggtaattg tgtcatttaa ggtcaacaag cagctgaaga
     1201 cggtcaacaa ctacttcctc ttaagcctgg cctgtgccga tctgattatc ggggtcattt
     1261 caatgaatct gtttacgacc tacatcatca tgaatcgatg ggccttaggg aacttggcct
     1321 gtgacctctg gcttgccatt gactgcgtag ccagcaatgc ctctgttatg aatcttctgg
     1381 tcatcagctt tgacagatac ttttccatca cgaggccgct cacgtaccga gccaaacgaa
     1441 caacaaagag agccggtgtg atgatcggtc tggcttgggt catctccttt gtcctttggg
     1501 ctcctgccat cttgttctgg caatactttg ttggaaagag aactgtgcct ccgggagagt
     1561 gcttcattca gttcctcagt gagcccacca ttacttttgg cacagccatc gctggttttt
     1621 atatgcctgt caccattatg actattttat actggaggat ctataaggaa actgaaaagc
     1681 gtaccaaaga gcttgctggc ctgcaagcct ctgggacaga ggcagagaca gaaaactttg
     1741 tccaccccac gggcagttct cgaagctgca gcagttacga acttcaacag caaagcatga
     1801 aacgctccaa caggaggaag tatggccgct gccacttctg gttcacaacc aagagctgga
     1861 aacccagctc cgagcagatg gaccaagacc acagcagcag tgacagttgg aacaacaatg
     1921 atgctgctgc ctccctggag aactccgcct cctccgacga ggaggacatt ggctccgaga
     1981 cgagagccat ctactccatc gtgctcaagc ttccgggtca cagcaccatc ctcaactcca
     2041 ccaagttacc ctcatcggac aacctgcagg tgcctgagga ggagctgggg atggtggact
     2101 tggagaggaa agccgacaag ctgcaggccc agaagagcgt ggacgatgga ggcagttttc
     2161 caaaaagctt ctccaagctt cccatccagc tagagtcagc cgtggacaca gctaagactt
     2221 ctgacgtcaa ctcctcagtg ggtaagagca cggccactct acctctgtcc ttcaaggaag
     2281 ccactctggc caagaggttt gctctgaaga ccagaagtca gatcactaag cggaaaagga
     2341 tgtccctggt caaggagaag aaagcggccc agaccctcag tgcgatcttg cttgccttca
     2401 tcatcacttg gaccccatac aacatcatgg ttctggtgaa caccttttgt gacagctgca
     2461 tacccaaaac cttttggaat ctgggctact ggctgtgcta catcaacagc accgtgaacc
     2521 ccgtgtgcta tgctctgtgc aacaaaacat tcagaaccac tttcaagatg ctgctgctgt
     2581 gccagtgtga caaaaaaaag aggcgcaagc agcagtacca gcagagacag tcggtcattt
     2641 ttcacaagcg cgcacccgag caggccttgt aggcggccgc gatatcgccc ctctccctcc
     2701 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2761 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2821 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2881 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2941 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3001 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3061 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3121 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3181 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3241 ctttgaaaaa cacgatgata agccaccatg gtgagcaagg gcgaggagct gttcaccggg
     3301 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3361 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     3421 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgatgtgc
     3481 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3541 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3601 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3661 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3721 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3781 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3841 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaaact gagcaaagac
     3901 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3961 ctcggcatgg acgagctgta caagtgatag gaattcgata tcaagcttat cgataatcaa
     4021 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4081 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4141 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4201 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4261 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4321 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4381 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     4441 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4501 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4561 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     4621 ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg
     4681 aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt
     4741 ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc
     4801 aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag
     4861 tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc
     4921 agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt
     4981 tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg
     5041 tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct
     5101 tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct
     5161 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     5221 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     5281 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5341 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5401 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5461 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5521 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5581 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5641 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5701 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5761 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     5821 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5881 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5941 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     6001 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     6061 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6121 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6181 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6241 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6301 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6361 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6421 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6481 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6541 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6601 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6661 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6721 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6781 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6841 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6901 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6961 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7021 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7081 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7141 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7201 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7261 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7321 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7381 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7441 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7501 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7561 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7621 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7681 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7741 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7801 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7861 gccttttgct ggccttttgc tcacatgt
//
