LOCUS       sequence-317858-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of  hM4Di-mCherry in a cre-dependent fashion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   Beier KT, Kim CK, Hoerbelt P, Hung LW, Heifets BD, DeLoach KE, Mosca
            TJ, Neuner S, Deisseroth K, Luo L, Malenka RC
  TITLE     Rabies screen reveals GPe control of cocaine-triggered plasticity.
  JOURNAL   Nature. 2017 Sep 21;549(7672):345-350. doi: 10.1038/nature23888. 
            Epub 2017 Sep 13.
  PUBMED    28902833
REFERENCE   2  (bases 1 to 7073)
  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"; doi: 
            "10.1038/nature23888"; journalName: "Nature"; date: "2017-09-21- 
            21"; volume: "549"; issue: "7672"; pages: "345-350"
COMMENT     Sequence Label: pAAV hSyn FLEx-loxp hM4Di-mCherry
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1459..1468
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1465..2175
                     /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(1610..1628)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1878..1897
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2182..2215)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2278..2311)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(2387..2834)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   2879..3008
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3070..3658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3150..3169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3829..4620,4621..4689))
                     /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     4452..4471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4690..4794)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4862..4880
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4918..4940)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5040..5059
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5076..5531)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5213..5234)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5425..5444
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5606..5746
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5617..5746
                     /label=AAV2 ITR
     polyA_signal    5786..6262
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6044..6063
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6294..6882
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6400..6411
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6809..6829
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    6930..6963
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    7034..7067
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
ORIGIN
        1 gccgccatgg ccaacttcac acctgtcaat ggcagctcgg gcaatcagtc cgtgcgcctg
       61 gtcacgtcat catcccacaa tcgctatgag acggtggaaa tggtcttcat tgccacagtg
      121 acaggctccc tgagcctggt gactgtcgtg ggcaacatcc tggtgatgct gtccatcaag
      181 gtcaacaggc agctgcagac agtcaacaac tacttcctct tcagcctggc gtgtgctgat
      241 ctcatcatag gcgccttctc catgaacctc tacaccgtgt acatcatcaa gggctactgg
      301 cccctgggcg ccgtggtctg cgacctgtgg ctggccctgg actgcgtggt gagcaacgcc
      361 tccgtcatga accttctcat catcagcttt gaccgctact tctgcgtcac caagcctctc
      421 acctaccctg cccggcgcac caccaagatg gcaggcctca tgattgctgc tgcctgggta
      481 ctgtccttcg tgctctgggc gcctgccatc ttgttctggc agtttgtggt gggtaagcgg
      541 acggtgcccg acaaccagtg cttcatccag ttcctgtcca acccagcagt gacctttggc
      601 acagccattg ctggcttcta cctgcctgtg gtcatcatga cggtgctgta catccacatc
      661 tccctggcca gtcgcagccg agtccacaag caccggcccg agggcccgaa ggagaagaaa
      721 gccaagacgc tggccttcct caagagccca ctaatgaagc agagcgtcaa gaagcccccg
      781 cccggggagg ccgcccggga ggagctgcgc aatggcaagc tggaggaggc ccccccgcca
      841 gcgctgccac cgccaccgcg ccccgtggct gataaggaca cttccaatga gtccagctca
      901 ggcagtgcca cccagaacac caaggaacgc ccagccacag agctgtccac cacagaggcc
      961 accacgcccg ccatgcccgc ccctcccctg cagccgcggg ccctcaaccc agcctccaga
     1021 tggtccaaga tccagattgt gacgaagcag acaggcaatg agtgtgtgac agccattgag
     1081 attgtgcctg ccacgccggc tggcatgcgc cctgcggcca acgtggcccg caagttcgcc
     1141 agcatcgctc gcaaccaggt gcgcaagaag cggcagatgg cggcccggga gcgcaaagtg
     1201 acacgaacga tctttgccat tctgctggcc ttcatcctca cctggacgcc ctacaacgtc
     1261 atggtcctgg tgaacacctt ctgccagagc tgcatccctg acacggtgtg gtccattggc
     1321 tactggctct gctacgtcaa cagcaccatc aaccctgcct gctatgctct gtgcaacgcc
     1381 acctttaaaa agaccttccg gcacctgctg ctgtgccagt atcggaacat cggcactgcc
     1441 aggcgggatc caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc
     1501 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     1561 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1621 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     1681 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     1741 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1801 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1861 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1921 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1981 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     2041 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     2101 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2161 gagctgtaca agtaagtcga cataacttcg tatagcatac attatacgaa gttatgtgtc
     2221 gatggtgatg cttggcaatt cgggtcgatg gtgaagcatt ggtcttccta ttttgatata
     2281 acttcgtata ggatacttta tacgaagtta tcccggccgg gcccgcggta tcaaatcaat
     2341 tctttgccaa aatgatgaga cagcacagga tccggtacct tctcgactgc gctctcaggc
     2401 acgacacgac tcctccgctg cccaccgcag actgaggcag cgctgagtcg ccggcgccgc
     2461 agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc ggtccggctg
     2521 ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag gggagtttgc gggggaccgg
     2581 cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg gcgcgcgccg ccaggcgggg
     2641 gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca cgcgcctcgc cgcatcctgt
     2701 ttcccctccc cctctctgat aggggatgcg caatttgggg aatgggggtt gggtgcttgt
     2761 ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc ccgcctcatc ctggtcctaa
     2821 aacccacttg cactcatacg cagggccctc tgcagtctag acacacgcgt gcggccgcag
     2881 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2941 gggcgaccaa aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3001 cctgcaggac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3781 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3841 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3901 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3961 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4021 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4081 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4141 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4201 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4261 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4321 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4381 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4441 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4501 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4561 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4621 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4681 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4741 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4801 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4861 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     4921 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4981 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     5041 ggcatcagag cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt
     5101 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg
     5161 caaaatccct tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg
     5221 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5281 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     5341 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5401 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct
     5461 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5521 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
     5581 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag
     5641 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     5701 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc
     5761 cgcacgtggt tacctacaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta
     5821 atcccagcaa tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca
     5881 gcctggccaa tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg
     5941 tcatgcatgc ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc
     6001 aggaggcgga gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat
     6061 agaccccgca ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc
     6121 tccaccccat aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta
     6181 ttaggacaag gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg
     6241 aggggtcaca gggatgccac ccgtagatct ctcgagcagc gctcggtatc gatgcgggga
     6301 ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg
     6361 cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta
     6421 gcagaaggac gtcccgcgca gaatccaggt ggcaacatag gcgagcagcc aaggaaagga
     6481 cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     6541 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa
     6601 agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc
     6661 aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga
     6721 cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     6781 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     6841 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa gcttgatatc
     6901 gaattcatcg atcggccgca tataagctta taacttcgta taatgtatgc tatacgaagt
     6961 tatgggtcga tggtgagatc tggactagag ggtcgatggt gatgcttgga tcaaaatagg
     7021 aagaatccca atgataactt cgtataaagt atcctatacg aagttatagg cgc
//
