LOCUS       sequence-360830-        7107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV to express hM4D(Gi)-mCherry fusion protein. Viral genome 3' UTR 
            modified to bring the viral genome below the optimal AAV packaging 
            limit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  TITLE     Ploski Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7107)
  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            "Unpublished"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-hM4D(Gi)-mCherry-WPRE-pA(Short)
FEATURES             Location/Qualifiers
     source          1..7107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   20..149
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        168..1456
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1489..1498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2947..2956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2953..3663
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3098..3116)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3366..3385
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3688..4276
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3741..3761)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4159..4170)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4331..4350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4389..4529
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4389..4518
                     /label=AAV2 ITR
     rep_origin      4604..5059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4691..4710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4901..4922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5076..5095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5195..5217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5255..5273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5341..5445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5446..5514,5515..6306)
                     /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(5664..5683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6477..7065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6966..6985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      121 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtttaacat tatggcctta
      181 ggtcacttca tctccatggg gttcttcttc tgattttcta gaaaatgaga tgggggtgca
      241 gagagcttcc tcagtgacct gcccagggtc acatcagaaa tgtcagagct agaacttgaa
      301 ctcagattac taatcttaaa ttccatgcct tgggggcatg caagtacgat atacagaagg
      361 agtgaactca ttagggcaga tgaccaatga gtttaggaaa gaagagtcca gggcagggta
      421 catctacacc acccgcccag ccctgggtga gtccagccac gttcacctca ttatagttgc
      481 ctctctccag tcctaccttg acgggaagca caagcagaaa ctgggacagg agccccagga
      541 gaccaaatct tcatggtccc tctgggagga tgggtgggga gagctgtggc agaggcctca
      601 ggaggggccc tgctgctcag tggtgacaga taggggtgag aaagcagaca gagtcattcc
      661 gtcagcattc tgggtctgtt tggtacttct tctcacgcta aggtggcggt gtgatatgca
      721 caatggctaa aaagcaggga gagctggaaa gaaacaagga cagagacaga ggccaagtca
      781 accagaccaa ttcccagagg aagcaaagaa accattacag agactacaag ggggaaggga
      841 aggagagatg aattagcttc ccctgtaaac cttagaaccc agctgttgcc agggcaacgg
      901 ggcaatacct gtctcttcag aggagatgaa gttgccaggg taactacatc ctgtctttct
      961 caaggaccat cccagaatgt ggcacccact agccgttacc atagcaactg cctctttgcc
     1021 ccacttaatc ccatcccgtc tgttaaaagg gccctatagt tggaggtggg ggaggtagga
     1081 agagcgatga tcacttgtgg actaagtttg ttcgcatccc cttctccaac cccctcagta
     1141 catcaccctg ggggaacagg gtccacttgc tcctgggccc acacagtcct gcagtattgt
     1201 gtatataagg ccagggcaaa gaggagcagg ttttaaagtg aaaggcaggc aggtgttggg
     1261 gaggcagtta ccggggcaac gggaacaggg cgtttcggag gtggttgcca tggggacctg
     1321 gatgctgacg aaggctcgcg aggctgtgag cagccacagt gccctgctca gaagccccaa
     1381 gctcgtcagt caagccggtt ctccgtttgc actcaggagc acgggcaggc gagtggcccc
     1441 tagttctggg ggcagctcta gagcggtacc ggatcctcta gagtcgacgc caccatggcc
     1501 aacttcacac ctgtcaatgg cagctcgggc aatcagtccg tgcgcctggt cacgtcatca
     1561 tcccacaatc gctatgagac ggtggaaatg gtcttcattg ccacagtgac aggctccctg
     1621 agcctggtga ctgtcgtggg caacatcctg gtgatgctgt ccatcaaggt caacaggcag
     1681 ctgcagacag tcaacaacta cttcctcttc agcctggcgt gtgctgatct catcataggc
     1741 gccttctcca tgaacctcta caccgtgtac atcatcaagg gctactggcc cctgggcgcc
     1801 gtggtctgcg acctgtggct ggccctggac tgcgtggtga gcaacgcctc cgtcatgaac
     1861 cttctcatca tcagctttga ccgctacttc tgcgtcacca agcctctcac ctaccctgcc
     1921 cggcgcacca ccaagatggc aggcctcatg attgctgctg cctgggtact gtccttcgtg
     1981 ctctgggcgc ctgccatctt gttctggcag tttgtggtgg gtaagcggac ggtgcccgac
     2041 aaccagtgct tcatccagtt cctgtccaac ccagcagtga cctttggcac agccattgct
     2101 ggcttctacc tgcctgtggt catcatgacg gtgctgtaca tccacatctc cctggccagt
     2161 cgcagccgag tccacaagca ccggcccgag ggcccgaagg agaagaaagc caagacgctg
     2221 gccttcctca agagcccact aatgaagcag agcgtcaaga agcccccgcc cggggaggcc
     2281 gcccgggagg agctgcgcaa tggcaagctg gaggaggccc ccccgccagc gctgccaccg
     2341 ccaccgcgcc ccgtggctga taaggacact tccaatgagt ccagctcagg cagtgccacc
     2401 cagaacacca aggaacgccc agccacagag ctgtccacca cagaggccac cacgcccgcc
     2461 atgcccgccc ctcccctgca gccgcgggcc ctcaacccag cctccagatg gtccaagatc
     2521 cagattgtga cgaagcagac aggcaatgag tgtgtgacag ccattgagat tgtgcctgcc
     2581 acgccggctg gcatgcgccc tgcggccaac gtggcccgca agttcgccag catcgctcgc
     2641 aaccaggtgc gcaagaagcg gcagatggcg gcccgggagc gcaaagtgac acgaacgatc
     2701 tttgccattc tgctggcctt catcctcacc tggacgccct acaacgtcat ggtcctggtg
     2761 aacaccttct gccagagctg catccctgac acggtgtggt ccattggcta ctggctctgc
     2821 tacgtcaaca gcaccatcaa ccctgcctgc tatgctctgt gcaacgccac ctttaaaaag
     2881 accttccggc acctgctgct gtgccagtat cggaacatcg gcactgccag gcgggatcca
     2941 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     3001 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     3061 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     3121 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3181 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3241 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3301 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3361 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3421 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3481 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3541 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3601 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3661 taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     3721 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3781 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3841 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3901 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3961 tccggaactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4021 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4081 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     4141 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4201 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4261 tgggccgcct ccccgcatcg ataccgagcg ctgctcgagt ttattgcagc ttataatggt
     4321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actggaccga
     4381 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4441 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4501 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4561 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4621 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4681 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4741 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4801 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4861 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4921 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4981 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5041 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5101 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5161 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5221 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5281 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5341 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5401 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5461 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5521 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5581 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5641 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5701 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5761 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5821 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5881 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5941 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6001 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6061 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6121 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6181 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6241 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6301 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6361 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6421 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6481 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6541 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6601 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6661 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6721 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6781 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6841 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6901 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6961 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7021 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7081 gcctttttac ggttcctggc cttttgc
//
