LOCUS       sequence-75495-e        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gs-coupled rM3D-IRES-mCitrine under the control of GFAP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7354)
  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            "Unpublished"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-rM3D(Gs)-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..7354
                     /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    2154..2727
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2321..2338)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2548..2567
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2728..2737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2734..2736,2737..2739,2740..3450)
                     /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(2779..2800)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3040..3059)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3387..3408
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3481..4069
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3534..3554)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3952..3963)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4101..4577
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4300..4319)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4617..4757
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4617..4746
                     /label=AAV2 ITR
     rep_origin      4832..5287
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4919..4938)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5129..5150
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5304..5323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5423..5445
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5483..5501)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5569..5673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5674..5742,5743..6534)
                     /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      6705..7293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7194..7213
                     /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 ctacgacgtc cccgactacg
      901 ccaccttgca cagtaacagt acaacctcgc ctttgtttcc caacatcagc tcttcctggg
      961 tgcacagtcc ctcggaggca gggctgccct tggggacagt cactcagttg ggcagctaca
     1021 acatttcaca agaaactggg aatttctcct caaacgacac ctccagcgac cctctcgggg
     1081 gtcacaccat ctggcaagtg gtcttcattg ccttcttaac tggcttcctg gcattggtga
     1141 ccatcattgg caacatcctt gtcattgtgg ccttcaaggt caacaaacag ctgaagacag
     1201 tcaacaacta cttcctctta agcctggcct gtgcagacct gatcatcggg gtcatttcca
     1261 tgaacctgtt cactacctac atcattatga accgttgggc actggggaac ttagcctgcg
     1321 acctctggct ctccattgac tgtgtggcca gcaatgcctc tgtcatgaat ctgctggtca
     1381 tcagctttga caggtacttt tccatcactt ctccattccg ctaccagagc ctgatgacca
     1441 gggctcgagc tggtgtgatg attggtctgg cttgggtcat ctcctttgtc ctatgggctc
     1501 ctgccatctt gttctggcaa tactttgtag ggaagagaac tgtgccccca ggagaatgtt
     1561 tcattcagtt tctgagtgag cccaccatca ccttcggcac ggcgatcgct ggcttttaca
     1621 tgcctgtcac catcatgact attttatact ggcgggtgta ccgggaggcc aaggagcaga
     1681 tcaggaagat cgaccgctgc gagggccggt tctatggcag ccaggagcag ccgcagccac
     1741 ccccgctccc ccaacaccag cccatcctcg gcaacggccg tgccagcaag aggaagacgt
     1801 cccgtgtcat ggccatgagg gaacacaaag ctctgcagac gctcagtgcc atcttgcttg
     1861 ccttcatcat cacgtggacc ccctacaaca tcatggtcct ggtgaacacc ttctgtgaca
     1921 gctgcatacc caaaacctat tggaatctgg gctactggct gtgctatatc aacagcaccg
     1981 tgaaccctgt gtgctatgcc ctgtgcaaca aaacattcag aaccaccttc aagacgctcc
     2041 tcttgtgcca gtgtgacaaa aggaagaggc gcaaacagca gtaccagcag agacagtcgg
     2101 tcatttttca caagcgagtg ccggagcagg ccttgtaggc ggccgcgata tcgcccctct
     2161 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     2221 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     2281 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     2341 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     2401 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     2461 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     2521 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     2581 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     2641 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     2701 gttttccttt gaaaaacacg atgataagcc accatggtga gcaagggcga ggagctgttc
     2761 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2821 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2881 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2941 atgtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3001 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3061 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3121 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3181 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3241 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3301 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caaactgagc
     3361 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3421 atcactctcg gcatggacga gctgtacaag tgataggaat tcgatatcaa gcttatcgat
     3481 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3541 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3601 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3661 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3721 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3781 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3841 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3901 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3961 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4021 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     4081 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     4141 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     4201 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     4261 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     4321 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     4381 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     4441 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     4501 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     4561 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     4621 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4681 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4741 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4801 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5041 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5161 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5281 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5341 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5401 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5461 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5521 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5581 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5641 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5701 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5761 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5821 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5881 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5941 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6001 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6061 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6121 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6181 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6241 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6301 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6361 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6421 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6481 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6541 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6601 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6661 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6721 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6781 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6841 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6901 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6961 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7021 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7081 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7141 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7201 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7261 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7321 ttcctggcct tttgctggcc ttttgctcac atgt
//
