LOCUS       sequence-305068-        8476 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 8476)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8476)
  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            "Unpublished"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..8476
                     /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        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             1467..1493
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3279..3852
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3446..3463)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3673..3692
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3853..3862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3859..3861,3862..3864,3865..4575)
                     /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(3904..3925)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4165..4184)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4512..4533
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4603..5191
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4656..4676)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5074..5085)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5223..5699
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5422..5441)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5739..5879
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5739..5868
                     /label=AAV2 ITR
     rep_origin      5954..6409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6041..6060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6251..6272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6426..6445)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6545..6567
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6691..6795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6796..6864,6865..7656)
                     /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(7014..7033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7827..8415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8316..8335
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccgcc accatgtacc catacgatgt tccagattac gctaccttgc
     1501 acaataacag tacaacctcg cctttgtttc caaacatcag ctcctcctgg atacacagcc
     1561 cctccgatgc agggctgccc ccgggaaccg tcactcattt cggcagctac aatgtttctc
     1621 gagcagctgg caatttctcc tctccagacg gtaccaccga tgaccctctg ggaggtcata
     1681 ccgtctggca agtggtcttc atcgctttct taacgggcat cctggccttg gtgaccatca
     1741 tcggcaacat cctggtaatt gtgtcattta aggtcaacaa gcagctgaag acggtcaaca
     1801 actacttcct cttaagcctg gcctgtgccg atctgattat cggggtcatt tcaatgaatc
     1861 tgtttacgac ctacatcatc atgaatcgat gggccttagg gaacttggcc tgtgacctct
     1921 ggcttgccat tgactgcgta gccagcaatg cctctgttat gaatcttctg gtcatcagct
     1981 ttgacagata cttttccatc acgaggccgc tcacgtaccg agccaaacga acaacaaaga
     2041 gagccggtgt gatgatcggt ctggcttggg tcatctcctt tgtcctttgg gctcctgcca
     2101 tcttgttctg gcaatacttt gttggaaaga gaactgtgcc tccgggagag tgcttcattc
     2161 agttcctcag tgagcccacc attacttttg gcacagccat cgctggtttt tatatgcctg
     2221 tcaccattat gactatttta tactggagga tctataagga aactgaaaag cgtaccaaag
     2281 agcttgctgg cctgcaagcc tctgggacag aggcagagac agaaaacttt gtccacccca
     2341 cgggcagttc tcgaagctgc agcagttacg aacttcaaca gcaaagcatg aaacgctcca
     2401 acaggaggaa gtatggccgc tgccacttct ggttcacaac caagagctgg aaacccagct
     2461 ccgagcagat ggaccaagac cacagcagca gtgacagttg gaacaacaat gatgctgctg
     2521 cctccctgga gaactccgcc tcctccgacg aggaggacat tggctccgag acgagagcca
     2581 tctactccat cgtgctcaag cttccgggtc acagcaccat cctcaactcc accaagttac
     2641 cctcatcgga caacctgcag gtgcctgagg aggagctggg gatggtggac ttggagagga
     2701 aagccgacaa gctgcaggcc cagaagagcg tggacgatgg aggcagtttt ccaaaaagct
     2761 tctccaagct tcccatccag ctagagtcag ccgtggacac agctaagact tctgacgtca
     2821 actcctcagt gggtaagagc acggccactc tacctctgtc cttcaaggaa gccactctgg
     2881 ccaagaggtt tgctctgaag accagaagtc agatcactaa gcggaaaagg atgtccctgg
     2941 tcaaggagaa gaaagcggcc cagaccctca gtgcgatctt gcttgccttc atcatcactt
     3001 ggaccccata caacatcatg gttctggtga acaccttttg tgacagctgc atacccaaaa
     3061 ccttttggaa tctgggctac tggctgtgct acatcaacag caccgtgaac cccgtgtgct
     3121 atgctctgtg caacaaaaca ttcagaacca ctttcaagat gctgctgctg tgccagtgtg
     3181 acaaaaaaaa gaggcgcaag cagcagtacc agcagagaca gtcggtcatt tttcacaagc
     3241 gcgcacccga gcaggccttg taggcggccg cgatatcgcc cctctccctc ccccccccct
     3301 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     3361 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3421 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3481 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3541 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3601 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3661 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3721 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3781 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3841 acacgatgat aagccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3901 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     3961 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4021 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc
     4081 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4141 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4201 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     4261 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     4321 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     4381 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4441 ctgctgcccg acaaccacta cctgagctac cagtccaaac tgagcaaaga ccccaacgag
     4501 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4561 gacgagctgt acaagtgaga attcgatatc aagcttatcg ataatcaacc tctggattac
     4621 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4681 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4741 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4801 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4861 acctgtcagc tcctttccgg aactttcgct ttccccctcc ctattgccac ggcggaactc
     4921 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4981 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     5041 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5101 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     5161 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     5221 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     5281 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     5341 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     5401 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     5461 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     5521 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     5581 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     5641 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     5701 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     5761 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     5821 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     5881 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5941 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6001 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     6061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     6181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     6301 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     6361 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6421 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6481 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6541 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6601 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6661 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6721 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6781 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6841 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6901 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6961 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7021 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7081 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7141 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7201 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7261 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7321 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7381 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7441 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7501 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7561 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7621 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7681 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7741 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7801 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7861 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7921 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7981 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8041 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8101 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8161 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8221 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8281 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8341 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8401 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8461 ccttttgctc acatgt
//
