LOCUS       sequence-75420-e        8490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gq-coupled hM3D-IRES-mCitrine under the control of CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8490)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8490)
  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            "Unpublished"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-CaMKIIa-HA-hM3D(Gq)-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..8490
                     /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
     promoter        160..1448
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             1478..1504
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3290..3863
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3457..3474)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3684..3703
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3864..3873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3870..3872,3873..3875,3876..4586)
                     /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(3915..3936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4176..4195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4523..4544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4617..5205
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4670..4690)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5088..5099)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5237..5713
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5436..5455)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5753..5893
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5753..5882
                     /label=AAV2 ITR
     rep_origin      5968..6423
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6055..6074)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6265..6286
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6440..6459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6559..6581
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6705..6809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6810..6878,6879..7670)
                     /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(7028..7047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7841..8429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8330..8349
                     /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 cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagctc tagagcggta ccggatccgc caccatgtac ccatacgatg ttccagatta
     1501 cgctaccttg cacaataaca gtacaacctc gcctttgttt ccaaacatca gctcctcctg
     1561 gatacacagc ccctccgatg cagggctgcc cccgggaacc gtcactcatt tcggcagcta
     1621 caatgtttct cgagcagctg gcaatttctc ctctccagac ggtaccaccg atgaccctct
     1681 gggaggtcat accgtctggc aagtggtctt catcgctttc ttaacgggca tcctggcctt
     1741 ggtgaccatc atcggcaaca tcctggtaat tgtgtcattt aaggtcaaca agcagctgaa
     1801 gacggtcaac aactacttcc tcttaagcct ggcctgtgcc gatctgatta tcggggtcat
     1861 ttcaatgaat ctgtttacga cctacatcat catgaatcga tgggccttag ggaacttggc
     1921 ctgtgacctc tggcttgcca ttgactgcgt agccagcaat gcctctgtta tgaatcttct
     1981 ggtcatcagc tttgacagat acttttccat cacgaggccg ctcacgtacc gagccaaacg
     2041 aacaacaaag agagccggtg tgatgatcgg tctggcttgg gtcatctcct ttgtcctttg
     2101 ggctcctgcc atcttgttct ggcaatactt tgttggaaag agaactgtgc ctccgggaga
     2161 gtgcttcatt cagttcctca gtgagcccac cattactttt ggcacagcca tcgctggttt
     2221 ttatatgcct gtcaccatta tgactatttt atactggagg atctataagg aaactgaaaa
     2281 gcgtaccaaa gagcttgctg gcctgcaagc ctctgggaca gaggcagaga cagaaaactt
     2341 tgtccacccc acgggcagtt ctcgaagctg cagcagttac gaacttcaac agcaaagcat
     2401 gaaacgctcc aacaggagga agtatggccg ctgccacttc tggttcacaa ccaagagctg
     2461 gaaacccagc tccgagcaga tggaccaaga ccacagcagc agtgacagtt ggaacaacaa
     2521 tgatgctgct gcctccctgg agaactccgc ctcctccgac gaggaggaca ttggctccga
     2581 gacgagagcc atctactcca tcgtgctcaa gcttccgggt cacagcacca tcctcaactc
     2641 caccaagtta ccctcatcgg acaacctgca ggtgcctgag gaggagctgg ggatggtgga
     2701 cttggagagg aaagccgaca agctgcaggc ccagaagagc gtggacgatg gaggcagttt
     2761 tccaaaaagc ttctccaagc ttcccatcca gctagagtca gccgtggaca cagctaagac
     2821 ttctgacgtc aactcctcag tgggtaagag cacggccact ctacctctgt ccttcaagga
     2881 agccactctg gccaagaggt ttgctctgaa gaccagaagt cagatcacta agcggaaaag
     2941 gatgtccctg gtcaaggaga agaaagcggc ccagaccctc agtgcgatct tgcttgcctt
     3001 catcatcact tggaccccat acaacatcat ggttctggtg aacacctttt gtgacagctg
     3061 catacccaaa accttttgga atctgggcta ctggctgtgc tacatcaaca gcaccgtgaa
     3121 ccccgtgtgc tatgctctgt gcaacaaaac attcagaacc actttcaaga tgctgctgct
     3181 gtgccagtgt gacaaaaaaa agaggcgcaa gcagcagtac cagcagagac agtcggtcat
     3241 ttttcacaag cgcgcacccg agcaggcctt gtaggcggcc gcgatatcgc ccctctccct
     3301 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     3361 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     3421 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     3481 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     3541 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     3601 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     3661 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     3721 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     3781 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     3841 tcctttgaaa aacacgatga taagccacca tggtgagcaa gggcgaggag ctgttcaccg
     3901 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3961 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     4021 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
     4081 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     4141 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4201 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4261 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4321 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4381 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4441 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccaaa ctgagcaaag
     4501 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4561 ctctcggcat ggacgagctg tacaagtgat aggaattcga tatcaagctt atcgataatc
     4621 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4681 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4741 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4801 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4861 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4921 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4981 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     5041 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5101 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5161 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     5221 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     5281 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     5341 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     5401 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     5461 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     5521 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     5581 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     5641 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     5701 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     5761 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     5821 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     5881 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5941 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     6001 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6061 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6121 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6181 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6241 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6301 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6361 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     6421 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     6481 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     6541 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     6601 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6661 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7201 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7681 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7981 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     8281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     8341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     8401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8461 tggccttttg ctggcctttt gctcacatgt
//
