LOCUS       sequence-469421-        7834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV2 vector containing Gi-coupled hM4D DREADD fused with mCherry 
            inside a double floxed (FLEX) cassette under the control of CBA 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7834)
  AUTHORS   Baccini G, Foggetti A, Wernet N, Engelhardt KA, Kronenbitter K, 
            Michels J, Kulik A, Wozny C, Wulff P
  TITLE     Parvalbumin expression identifies subicular principal cells with 
            high projection specificity
  JOURNAL   bioRxiv 2024.08.13.606964
REFERENCE   2  (bases 1 to 7834)
  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: "bioRxiv 
            2024.08.13.606964"
COMMENT     Sequence Label: pAM-FLEX-hM4D(Gi)-mCherry
FEATURES             Location/Qualifiers
     source          1..7834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     promoter        442..719
                     /label=chicken beta-actin promoter
     primer_bind     998..1020
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1082..1101
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1136..1169
                     /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)."
     protein_bind    1232..1265
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1277..1987)
                     /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(1555..1574)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1824..1842
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1984..1993
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3442..3451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3484..3517)
                     /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)."
     protein_bind    complement(3588..3621)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3660..4248
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3713..3733)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4131..4142)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4275..4292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4281..4505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4715..5044)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4729..4864
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5029..5049
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5290..5878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5370..5389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6049..6840,6841..6909))
                     /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     6672..6691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6910..7014)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7082..7100
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7138..7160)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7260..7279
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7493..7512
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7802..7820
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7802..7819
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      481 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      541 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      601 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      661 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      721 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      781 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      841 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      901 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
      961 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1021 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tccccataac
     1141 ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg cttcaccatc
     1201 gacccgaatt gccaagcatc accatcgacc cataacttcg tataatgtat gctatacgaa
     1261 gttatgaatt cgcgccttac ttgtacagct cgtccatgcc gccggtggag tggcggccct
     1321 cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccaact
     1381 tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg taggtggtct
     1441 tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg atctcgccct
     1501 tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc atggtcttct
     1561 tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga
     1621 actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg tcctcgaagt
     1681 tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt
     1741 cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac aggatgtccc
     1801 aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg gtgccctcgt
     1861 aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca
     1921 tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc tcgcccttgc
     1981 tcaccatggt ggcgaccggt ggatcccgcc tggcagtgcc gatgttccga tactggcaca
     2041 gcagcaggtg ccggaaggtc tttttaaagg tggcgttgca cagagcatag caggcagggt
     2101 tgatggtgct gttgacgtag cagagccagt agccaatgga ccacaccgtg tcagggatgc
     2161 agctctggca gaaggtgttc accaggacca tgacgttgta gggcgtccag gtgaggatga
     2221 aggccagcag aatggcaaag atcgttcgtg tcactttgcg ctcccgggcc gccatctgcc
     2281 gcttcttgcg cacctggttg cgagcgatgc tggcgaactt gcgggccacg ttggccgcag
     2341 ggcgcatgcc agccggcgtg gcaggcacaa tctcaatggc tgtcacacac tcattgcctg
     2401 tctgcttcgt cacaatctgg atcttggacc atctggaggc tgggttgagg gcccgcggct
     2461 gcaggggagg ggcgggcatg gcgggcgtgg tggcctctgt ggtggacagc tctgtggctg
     2521 ggcgttcctt ggtgttctgg gtggcactgc ctgagctgga ctcattggaa gtgtccttat
     2581 cagccacggg gcgcggtggc ggtggcagcg ctggcggggg ggcctcctcc agcttgccat
     2641 tgcgcagctc ctcccgggcg gcctccccgg gcgggggctt cttgacgctc tgcttcatta
     2701 gtgggctctt gaggaaggcc agcgtcttgg ctttcttctc cttcgggccc tcgggccggt
     2761 gcttgtggac tcggctgcga ctggccaggg agatgtggat gtacagcacc gtcatgatga
     2821 ccacaggcag gtagaagcca gcaatggctg tgccaaaggt cactgctggg ttggacagga
     2881 actggatgaa gcactggttg tcgggcaccg tccgcttacc caccacaaac tgccagaaca
     2941 agatggcagg cgcccagagc acgaaggaca gtacccaggc agcagcaatc atgaggcctg
     3001 ccatcttggt ggtgcgccgg gcagggtagg tgagaggctt ggtgacgcag aagtagcggt
     3061 caaagctgat gatgagaagg ttcatgacgg aggcgttgct caccacgcag tccagggcca
     3121 gccacaggtc gcagaccacg gcgcccaggg gccagtagcc cttgatgatg tacacggtgt
     3181 agaggttcat ggagaaggcg cctatgatga gatcagcaca cgccaggctg aagaggaagt
     3241 agttgttgac tgtctgcagc tgcctgttga ccttgatgga cagcatcacc aggatgttgc
     3301 ccacgacagt caccaggctc agggagcctg tcactgtggc aatgaagacc atttccaccg
     3361 tctcatagcg attgtgggat gatgacgtga ccaggcgcac ggactgattg cccgagctgc
     3421 cattgacagg tgtgaagttg gccatggtgg cgctagtgcg gccgcctcga gagatctgat
     3481 atcataactt cgtataggat actttatacg aagttatcat tgggattctt cctattttga
     3541 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata
     3601 gcatacatta tacgaagtta tgtccctcga agaggttcaa gctttaacag cttatcgata
     3661 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3721 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3781 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3841 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3901 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3961 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4021 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4081 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4141 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4201 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     4261 gactcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4321 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4381 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4441 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4501 tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcatggc tacgtagata
     4561 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     4621 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     4681 gcggcctcag tgagcgagcg agcgcgcaga gctttttgca aaagcctagg cctccaaaaa
     4741 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     4801 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     4861 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     4921 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     4981 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     5041 acacattcca cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5101 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5161 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5221 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5281 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5341 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5401 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5461 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5521 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5581 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5641 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5701 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5761 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5821 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5881 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5941 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6001 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6061 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6121 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6181 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6241 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6301 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6361 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6421 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6481 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6541 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6601 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6661 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6721 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6781 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6841 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6901 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6961 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7021 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7081 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     7141 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7201 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7261 ggcatcagag cagattgtac tgagagtgca ccattcgacg ctctccctta tgcgactcct
     7321 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     7381 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     7441 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     7501 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     7561 cggcgtagag gatctggcta gcgatgaccc tgctgattgg ttcgctgacc atttccgggt
     7621 gcgggacggc gttaccagaa actcagaagg ttcgtccaac caaaccgact ctgacggcag
     7681 tttacgagag agatgatagg gtctgcttca gtaagccaga tgctacacaa ttaggcttgt
     7741 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     7801 gatttaggtg acactataga atacacggaa ttaa
//
