LOCUS       sequence-75484-e        7953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gs-coupled rM3D-IRES-mCitrine under the control of CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7953)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7953)
  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            "Unpublished"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-rM3D(Gs)-IRES-mCitrine
FEATURES             Location/Qualifiers
     source          1..7953
                     /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    2756..3329
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2923..2940)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3150..3169
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3330..3339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3336..3338,3339..3341,3342..4052)
                     /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(3381..3402)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3642..3661)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3989..4010
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4080..4668
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4133..4153)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4551..4562)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4700..5176
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4899..4918)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5216..5356
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5216..5345
                     /label=AAV2 ITR
     rep_origin      5431..5886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5518..5537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5728..5749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5903..5922)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6022..6044
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6082..6100)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6168..6272
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6273..6341,6342..7133)
                     /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(6491..6510)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7304..7892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7793..7812
                     /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 ccctacgacg tccccgacta
     1501 cgccaccttg cacagtaaca gtacaacctc gcctttgttt cccaacatca gctcttcctg
     1561 ggtgcacagt ccctcggagg cagggctgcc cttggggaca gtcactcagt tgggcagcta
     1621 caacatttca caagaaactg ggaatttctc ctcaaacgac acctccagcg accctctcgg
     1681 gggtcacacc atctggcaag tggtcttcat tgccttctta actggcttcc tggcattggt
     1741 gaccatcatt ggcaacatcc ttgtcattgt ggccttcaag gtcaacaaac agctgaagac
     1801 agtcaacaac tacttcctct taagcctggc ctgtgcagac ctgatcatcg gggtcatttc
     1861 catgaacctg ttcactacct acatcattat gaaccgttgg gcactgggga acttagcctg
     1921 cgacctctgg ctctccattg actgtgtggc cagcaatgcc tctgtcatga atctgctggt
     1981 catcagcttt gacaggtact tttccatcac ttctccattc cgctaccaga gcctgatgac
     2041 cagggctcga gctggtgtga tgattggtct ggcttgggtc atctcctttg tcctatgggc
     2101 tcctgccatc ttgttctggc aatactttgt agggaagaga actgtgcccc caggagaatg
     2161 tttcattcag tttctgagtg agcccaccat caccttcggc acggcgatcg ctggctttta
     2221 catgcctgtc accatcatga ctattttata ctggcgggtg taccgggagg ccaaggagca
     2281 gatcaggaag atcgaccgct gcgagggccg gttctatggc agccaggagc agccgcagcc
     2341 acccccgctc ccccaacacc agcccatcct cggcaacggc cgtgccagca agaggaagac
     2401 gtcccgtgtc atggccatga gggaacacaa agctctgcag acgctcagtg ccatcttgct
     2461 tgccttcatc atcacgtgga ccccctacaa catcatggtc ctggtgaaca ccttctgtga
     2521 cagctgcata cccaaaacct attggaatct gggctactgg ctgtgctata tcaacagcac
     2581 cgtgaaccct gtgtgctatg ccctgtgcaa caaaacattc agaaccacct tcaagacgct
     2641 cctcttgtgc cagtgtgaca aaaggaagag gcgcaaacag cagtaccagc agagacagtc
     2701 ggtcattttt cacaagcgag tgccggagca ggccttgtag gcggccgcga tatcgcccct
     2761 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2821 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2881 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2941 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     3001 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     3061 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3121 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3181 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3241 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3301 tggttttcct ttgaaaaaca cgatgataag ccaccatggt gagcaagggc gaggagctgt
     3361 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3421 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     3481 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc
     3541 tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     3601 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     3661 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     3721 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     3781 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     3841 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     3901 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaaactga
     3961 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     4021 ggatcactct cggcatggac gagctgtaca agtgagaatt cgatatcaag cttatcgata
     4081 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4141 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4201 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4261 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4321 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4381 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4441 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4501 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4561 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4621 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     4681 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4741 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4801 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4861 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4921 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4981 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     5041 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     5101 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     5161 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     5221 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     5281 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     5341 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5401 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     5461 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5521 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5581 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5641 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5701 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5761 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5821 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5881 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5941 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6001 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6061 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6121 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6181 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6241 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6301 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6361 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6421 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6481 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6541 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6601 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6661 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6721 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6781 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6841 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6901 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6961 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7021 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7081 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7141 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7201 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7261 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7321 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7381 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7441 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7501 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7561 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7621 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7681 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7741 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7801 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7861 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7921 tcctggcctt ttgctggcct tttgctcaca tgt
//
