LOCUS       sequence-487712-        6852 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  AAV vector for cocaine-gated chemogenetic cation channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6852)
  AUTHORS   Gomez JL, Magnus CJ, Bonaventura J, Solis O, Curry FP, Levinstein 
            MR, Budinich RC, Carlton ML, Ventriglia EN, Lam S, Wang L, 
            Schoenborn I, Dunne W, Michaelides M, Sternson SM
  TITLE     Cocaine chemogenetics blunts drug-seeking by synthetic physiology.
  JOURNAL   Nature. 2025 Aug 27. doi: 10.1038/s41586-025-09427-8.
  PUBMED    40866713
REFERENCE   2  (bases 1 to 6852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2025 Aug 27. doi: 10.1038/s41586-025-09427-8."
COMMENT     Sequence Label: AAV Syn-coca-5HT3-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..6852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   826..955
                     /label=AAV2 ITR
     polyA_signal    1117..1237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1207..1226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1428..1448
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1521..2228)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1799..1818)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2068..2086
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2252..2803
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2254..2820
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2642..2659
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(4308..4327)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4310..4442
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   5035..5164
                     /label=AAV2 ITR
     rep_origin      5236..5691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5323..5342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5533..5554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5717..5821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5822..5890,5891..6682)
                     /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(6040..6059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      961 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccttatc
     1021 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca tctccccctg
     1081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1141 ggttacaaat aaagcaatag catcacaaat tcacaaataa agcatttttt tcactgcatt
     1201 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg aagcgcacca
     1261 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     1321 tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca
     1381 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     1441 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     1501 tactcgagac gcgtaccggt ttacttgtac agctcgtcca tgccgccggt ggagtggcgg
     1561 ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct cgttgtggga ggtgatgtcc
     1621 aacttgatgt tgacgttgta ggcgccgggc agctgcacgg gcttcttggc cttgtaggtg
     1681 gtcttgacct cagcgtcgta gtggccgccg tccttcagct tcagcctctg cttgatctcg
     1741 cccttcaggg cgccgtcctc ggggtacatc cgctcggagg aggcctccca gcccatggtc
     1801 ttcttctgca ttacggggcc gtcggagggg aagttggtgc cgcgcagctt caccttgtag
     1861 atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccacgcc gccgtcctcg
     1921 aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttcaa gtagtcgggg
     1981 atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca tgaactgagg ggacaggatg
     2041 tcccaggcga agggcagggg gccacccttg gtcaccttca gcttggcggt ctgggtgccc
     2101 tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt cacggagccc
     2161 tccatgtgca ccttgaagcg catgaactcc ttgatgatgg ccatgttatc ctcctcgccc
     2221 ttgctcaccg gatcccccat ggttgtggcc atattatcat cgtgtttttc aaaggaaaac
     2281 cacgtccccg tggttcgggg ggcctagacg tttttttaac ctcgactaaa cacatgtaaa
     2341 gcatgtgcac cgaggcccca gatcagatcc catacaatgg ggtaccttct gggcatcctt
     2401 cagccccttg ttgaatacgc ttaggagagc catttgactc tttccacaac tatccaactc
     2461 acaacgtggc actggggttg tgccgccttt gcaggtgtat cttatacacg tggcttttgg
     2521 ccgcagaggc acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt cgctacagac
     2581 gttgtttgtc ttcaagaagc ttccagagga actgcttcct tcacgacatt caacagacct
     2641 tgcattcctt tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa gacagggcca
     2701 ggtttccggg ccctcacatt gccaaaagac ggcaatatgg tggaaaataa catatagaca
     2761 aacgcacacc ggccttattc caagcggctt cggccagtaa cgttaggggg gggggrggga
     2821 grgggggggk ggggggrgks ggggggaatt gcggccgcct aactgtagtg ccatatggac
     2881 cagagtgtca ccagagtaat ggaataagcc agcaccgcca gcaagtagat ccgaaacagc
     2941 agcctgtcaa ggacgtagcc cactcgcagc cagtcgcgag cgacttcgcg catctcgtcc
     3001 cgcttctcca gaaaatgtcg gatgcttgac agttcctgga gcagcccccg gacagcgaga
     3061 gaggcttctc ggggaggagg aaggggtgat ccccgccccc gtggggtctt ttccaaatct
     3121 tgtggtcccc caacatggga gcagtgattc cccatagcag gcaacagatc agacccacta
     3181 caatcgtctg tcttgttggc ttgaaaggtt gcaggtggcc ggtgggccat aggttgctcg
     3241 ccaagacaca gaatccatgc aattctgtcc agcacaaggt gtcgcaacca gtctggcacg
     3301 ggtctttgca ggtcctgctt atgtaccagc cgtacaatga agatagtttc cgccagactt
     3361 atcaccagga gtgccataca cacgacgaaa tacactccta tcagcggagt gccgattgtg
     3421 gcaggaaggg tatcggacac gatgatcaga aatacagaat agccgagcaa caaggtaatc
     3481 ttaaaggaga ccctctcccc gctatcaggt gggagacaaa atcccacgat gtccacgacc
     3541 atgagaaaaa tactggggag caacaaacta acggcgtaga acaatggcct tcttctgatg
     3601 ataactgtga atgttacgtc agggaagggt tccttgcagc attcaaaaaa tcgctcgctt
     3661 cttttaccgg ggattcccac aaggtcccac tccccattcg ggatgtaacc agagatgtca
     3721 gcctcctgca tttgcagatc caggctccat ttgccgtagc tccagctccc gaatttgagt
     3781 ttgcagtgct ggacgtcgaa gggaaaccag cggacgtcga tatagcatga ggacttgaag
     3841 attccagggg gtccgtactg gcagtggcct gaagaattga ccaacacgtt ggtgtgaaat
     3901 gttgcatcaa acctttcgtc tgcgctgttg tacagcagga tatctggttt ccagatttgg
     3961 ccgtcaggga atcttacagt ctttacacct gggtattcgc tcacattcca ctgcaagtag
     4021 tgatcggtcc aggacatctg caaccaaata ttagtggtca gcacctggtt cttttcatcc
     4081 acgtccatga tctgcagcaa ggagaggctg aagtacactg tcagtggctg ggaatcattt
     4141 gcgaccggcc gctccagggg gttataattc ttaacaagct ccttatacag tttgcgctgg
     4201 aactcaccct gcaggctaac gtgcagaagg gaagcagcca gggcgagcca cacgccgcct
     4261 ggagaacagc gcatggtggc gaattcgcta gcactagtga tatcatcacc tgtggagaga
     4321 aaggcaaagt ggatgtcagt aagaccaata ggtgcctatc agaaacgcaa gagtcttctc
     4381 tgtctcgaca agcccagttt ctattggtct ccttaaacct gtcttgtaac cttgatactt
     4441 acctgatcca acggctgcga cttggggcag ggggtcctag gggtggtctg gccaggagcc
     4501 gcgggggcgg actgcgcggt gcccaggagc acagctgcgc tctcaggcac gacacgactc
     4561 ctccgctgcc caccgcagac tgaggcagcg ctgagtcgcc ggcgccgcag cgcagatggt
     4621 cgcgcccgtg cccccctatc tcgcgcctcg cgtggtgcgg tccggctggg ccggcggcgg
     4681 cgcggacgcg accaaggtgg ccgggaaggg gagtttgcgg gggaccggcg agtgacgtca
     4741 gcgcgccttc agtgctgagg cggcggtggc gcgcgccgcc aggcgggggc gaaggcactg
     4801 tccgcggtgc tgaagctggc agtgcgcacg cgcctcgccg catcctgttt cccctccccc
     4861 tctctgatag gggatgcgca atttggggaa tgggggttgg gtgcttgtcc agtgggtcgg
     4921 ggtcggtcgt caatttaaat actgaaaagg gcgaattcca gcacactgga gctcggatct
     4981 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     5041 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     5101 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     5161 gcagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5221 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5281 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5581 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5641 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     5701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5821 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5881 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5941 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6001 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6061 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6121 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6181 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6241 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6301 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6361 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6421 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6481 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6541 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6601 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6661 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6721 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6781 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6841 caaaggatct tc
//
