LOCUS       sequence-317852-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of  hM4Di-mCherry in a Flp-dependent fashion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  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 7126)
  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-FRT hM4Di-mCherry
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..48
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      1509..1518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1515..2225
                     /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(1660..1678)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1928..1947
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2341..2388)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(2466..2913)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   2958..3087
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3149..3737)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3229..3248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3908..4699,4700..4768))
                     /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     4531..4550
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4769..4873)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4941..4959
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4997..5019)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5119..5138
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5155..5610)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5292..5313)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5504..5523
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5685..5825
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5696..5825
                     /label=AAV2 ITR
     polyA_signal    5865..6341
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6123..6142
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6373..6961
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6479..6490
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6888..6908
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcgg cgcgccatgg
       61 ccaacttcac acctgtcaat ggcagctcgg gcaatcagtc cgtgcgcctg gtcacgtcat
      121 catcccacaa tcgctatgag acggtggaaa tggtcttcat tgccacagtg acaggctccc
      181 tgagcctggt gactgtcgtg ggcaacatcc tggtgatgct gtccatcaag gtcaacaggc
      241 agctgcagac agtcaacaac tacttcctct tcagcctggc gtgtgctgat ctcatcatag
      301 gcgccttctc catgaacctc tacaccgtgt acatcatcaa gggctactgg cccctgggcg
      361 ccgtggtctg cgacctgtgg ctggccctgg actgcgtggt gagcaacgcc tccgtcatga
      421 accttctcat catcagcttt gaccgctact tctgcgtcac caagcctctc acctaccctg
      481 cccggcgcac caccaagatg gcaggcctca tgattgctgc tgcctgggta ctgtccttcg
      541 tgctctgggc gcctgccatc ttgttctggc agtttgtggt gggtaagcgg acggtgcccg
      601 acaaccagtg cttcatccag ttcctgtcca acccagcagt gacctttggc acagccattg
      661 ctggcttcta cctgcctgtg gtcatcatga cggtgctgta catccacatc tccctggcca
      721 gtcgcagccg agtccacaag caccggcccg agggcccgaa ggagaagaaa gccaagacgc
      781 tggccttcct caagagccca ctaatgaagc agagcgtcaa gaagcccccg cccggggagg
      841 ccgcccggga ggagctgcgc aatggcaagc tggaggaggc ccccccgcca gcgctgccac
      901 cgccaccgcg ccccgtggct gataaggaca cttccaatga gtccagctca ggcagtgcca
      961 cccagaacac caaggaacgc ccagccacag agctgtccac cacagaggcc accacgcccg
     1021 ccatgcccgc ccctcccctg cagccgcggg ccctcaaccc agcctccaga tggtccaaga
     1081 tccagattgt gacgaagcag acaggcaatg agtgtgtgac agccattgag attgtgcctg
     1141 ccacgccggc tggcatgcgc cctgcggcca acgtggcccg caagttcgcc agcatcgctc
     1201 gcaaccaggt gcgcaagaag cggcagatgg cggcccggga gcgcaaagtg acacgaacga
     1261 tctttgccat tctgctggcc ttcatcctca cctggacgcc ctacaacgtc atggtcctgg
     1321 tgaacacctt ctgccagagc tgcatccctg acacggtgtg gtccattggc tactggctct
     1381 gctacgtcaa cagcaccatc aaccctgcct gctatgctct gtgcaacgcc acctttaaaa
     1441 agaccttccg gcacctgctg ctgtgccagt atcggaacat cggcactgcc aggcgggatc
     1501 caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     1561 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     1621 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     1681 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     1741 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     1801 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     1861 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     1921 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     1981 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     2041 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     2101 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     2161 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     2221 agtaagtcga cgaagttcct ataccttttg aagaatagga acttcggaat aggaacttct
     2281 gtcgatggtg atgcttggca attcgggtcg atggtgaagc attggtcttc ctattttgat
     2341 gaagttccta tactttctag agaataggaa cttcggaata ggaacttcgg tacccccggg
     2401 cccgcggtat caaatcaatt ctttgccaaa atgatgagac agcacaggat ccggtacctt
     2461 ctcgactgcg ctctcaggca cgacacgact cctccgctgc ccaccgcaga ctgaggcagc
     2521 gctgagtcgc cggcgccgca gcgcagatgg tcgcgcccgt gcccccctat ctcgcgcctc
     2581 gcgtggtgcg gtccggctgg gccggcggcg gcgcggacgc gaccaaggtg gccgggaagg
     2641 ggagtttgcg ggggaccggc gagtgacgtc agcgcgcctt cagtgctgag gcggcggtgg
     2701 cgcgcgccgc caggcggggg cgaaggcact gtccgcggtg ctgaagctgg cagtgcgcac
     2761 gcgcctcgcc gcatcctgtt tcccctcccc ctctctgata ggggatgcgc aatttgggga
     2821 atgggggttg ggtgcttgtc cagtgggtcg gggtcggtcg tcaggtaggc acccccaccc
     2881 cgcctcatcc tggtcctaaa acccacttgc actcatacgc agggccctct gcagtctaga
     2941 cacacgcgtg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3001 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga
     3061 gcgagcgagc gcgcagctgc ctgcaggaca tgtgagcaaa aggccagcaa aaggccagga
     3121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3301 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3361 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3421 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3481 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3541 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3601 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3661 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3721 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3901 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3961 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4021 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4081 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4141 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4201 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4261 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4321 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4381 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4441 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4501 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4561 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4621 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4681 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4741 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4801 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4861 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4921 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     4981 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5041 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5101 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     5161 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5221 accaatagac cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagagt
     5281 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5341 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     5401 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5461 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5521 gagcgggcgc taaggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5581 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     5641 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgcccctgca ggcagctgcg
     5701 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     5761 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     5821 ttcctgcggc cgctcggtcc gcacgtggtt acctacaaaa tcagaaggac agggaaggga
     5881 gcagtggttc acgcctgtaa tcccagcaat ttgggaggcc aaggtgggta gatcacctga
     5941 gattaggagt tggagaccag cctggccaat atggtgaaac cccgtctcta ccaaaaaaac
     6001 aaaaattagc tgagcctggt catgcatgcc tggaatccca acaactcggg aggctgaggc
     6061 aggagaatcg cttgaaccca ggaggcggag attgcagtga gccaagattg tgccactgca
     6121 ctccagcttg gttcccaata gaccccgcag gccctacagg ttgtcttccc aacttgcccc
     6181 ttgctccata ccacccccct ccaccccata atattataga aggacaccta gtcagacaaa
     6241 atgatgcaac ttaattttat taggacaagg ctggtgggca ctggagtggc aacttccagg
     6301 gccaggagag gcactgggga ggggtcacag ggatgccacc cgtagatctc tcgagcagcg
     6361 ctcggtatcg atgcggggag gcggcccaaa gggagatccg actcgtctga gggcgaaggc
     6421 gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg tccgctggat
     6481 tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg gcaacatagg
     6541 cgagcagcca aggaaaggac gatgatttcc ccgacaacac cacggaattg tcagtgccca
     6601 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     6661 tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca atgccccaac
     6721 cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg acaacgggcc
     6781 acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa atgaaagcca
     6841 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
     6901 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
     6961 tatcgataag cttgatatcg aattcatcga tcggccgcat ataagcttga agttcctatt
     7021 ccgaagttcc tattcttcaa aaggtatagg aacttcgggt cgatggtgag atctggacta
     7081 gagggtcgat ggtgatgctt ggatcaaaat aggaagaatc ccaatg
//
