LOCUS       sequence-432193-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides AND intersectional (Cre+Flp) expression of HA-hM4Di.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  AUTHORS   Hughes AC, Pollard BG, Xu B, Gammons JW, Chapman P, Bikoff JB, 
            Schwarz LA
  TITLE     A Novel Single Vector Intersectional AAV Strategy for Interrogating 
            Cellular Diversity and Brain Function.
  JOURNAL   bioRxiv. 2023 Feb 8:2023.02.07.527312. doi: 
            10.1101/2023.02.07.527312. Preprint.
  PUBMED    36798174
REFERENCE   2  (bases 1 to 5891)
  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. 
            2023 Feb 8:2023.02.07.527312. doi: 10.1101/2023.02.07.527312. 
            Preprint."
COMMENT     Sequence Label: pAAV-hSyn-ConVERGD-(HA)hM4Di-W3SL
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      928..1516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1417..1436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1840..1845,1846..1863,1864..1870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1878..1894
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1883..1905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1902..1918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1958..2098
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1969..2098
                     /label=AAV2 ITR
     primer_bind     2471..2491
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2595..2642
                     /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)."
     CDS             2761..2787
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    4229..4262
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4355..4388
                     /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(4401..4448)
                     /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(4557..5004)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   5038..5167
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     complement(5193..5210)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5193..5209)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5202..5224)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5418..5437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5537..5559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5597..5615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5683..5787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5788..5856,5857..5891,1..757)
                     /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"
ORIGIN
        1 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt ggttctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagctcggcg cgccattggg atccaatcct gcaggcagct gcgcgctcgc
     1981 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     2041 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgg
     2101 agctccggtt accggccgct ttaaaaaacc tcccacatct ccccctgaac ctgaaacata
     2161 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2221 gcaatagcat cacaaatttc acaaataaag ctagatggtt acaaataaag caatagcatc
     2281 acaaatttca caaataaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     2341 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca agaactaacc aggatttata
     2401 caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag gcattaaagc
     2461 agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca atctttcaca
     2521 aattttgtaa tccagaggtt gattatctcg ggcatcgaat atcgaatcgg ctaagctcta
     2581 gaaagcttcg tacggaagtt actattccga agttcctatt ctctagaaag tataggaact
     2641 tcgtgtcgat ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt cttcctattt
     2701 tgatgaagtt cctattcttc aaaaggtata ggaacttcct cgagaccggt agccaccatg
     2761 tacccatacg atgttccaga ttacgctatg gccaacttca caccagtcaa tggcagctcg
     2821 ggcaatcagt ccgtgcgcct ggtcacgtca tcatcccaca atcgctatga gacggtggaa
     2881 atggtcttca ttgccacagt gacaggctcc ctgagcctgg tgactgtcgt gggcaacatc
     2941 ctggtgatgc tgtccatcaa ggtcaacagg cagctgcaga cagtcaacaa ctacttcctc
     3001 ttcagcctgg cgtgtgctga tctcatcata ggcgccttct ccatgaacct ctacaccgtg
     3061 tacatcatca agggctactg gcccctgggc gccgtggtct gcgacctgtg gctggccctg
     3121 gactgcgtgg tgagcaacgc ctccgtcatg aaccttctca tcatcagctt tgaccgctac
     3181 ttctgcgtca ccaagcctct cacctaccct gcccggcgca ccaccaagat ggcaggcctc
     3241 atgattgctg ctgcctgggt actgtccttc gtgctctggg cgcctgccat cttgttctgg
     3301 cagtttgtgg tgggtaagcg gacggtgccc gacaaccagt gcttcatcca gttcctgtcc
     3361 aacccagcag tgacctttgg cacagccatt gctggcttct acctgcctgt ggtcatcatg
     3421 acggtgctgt acatccacat ctccctggcc agtcgcagcc gagtccacaa gcaccggccc
     3481 gagggcccga aggagaagaa agccaagacg ctggccttcc tcaagagccc actaatgaag
     3541 cagagcgtca agaagccccc gcccggggag gccgcccggg aggagctgcg caatggcaag
     3601 ctggaggagg cccccccgcc agcgctgcca ccgccaccgc gccccgtggc tgataaggac
     3661 acttccaatg agtccagctc aggcagtgcc acccagaaca ccaaggaacg cccagccaca
     3721 gagctgtcca ccacagaggc caccacgccc gccatgcccg cccctcccct gcagccgcgg
     3781 gccctcaacc cagcctccag atggtccaag atccagattg tgacgaagca gacaggcaat
     3841 gagtgtgtga cagccattga gattgtgcct gccacgccgg ctggcatgcg ccctgcggcc
     3901 aacgtggccc gcaagttcgc cagcatcgct cgcaaccagg tgcgcaagaa gcggcagatg
     3961 gcggcccggg agcgcaaagt gacacgaacg atctttgcca ttctgcttgc cttcatcctc
     4021 acctggacgc cctacaacgt catggtcctg gtgaacacct tctgccagag ctgcatccct
     4081 gacacggtgt ggtccattgg ctactggctc tgctacgtca acagcaccat caaccctgcc
     4141 tgctatgctc tgtgcaacgc cacctttaaa aagaccttcc ggcacctgct gctgtgccag
     4201 tatcggaaca tcggcactgc caggtagcat aacttcgtat aatgtatgct atacgaagtt
     4261 attttctttc tttgcgcgtc ctggattcgc ggaaacgcgt acatccagct gacgagtccc
     4321 aaataggacg aaacgcgctt ttctttttgt ttttataact tcgtataatg tatgctatac
     4381 gaagttatct taaggtcgac gaagttccta tactttctag agaataggaa cttcggaata
     4441 ggaacttcgt gtcgatggtg atgcttggca attcgggtcg atggtgaagc attggtcttc
     4501 ctattttgat gaagttccta taccttttga agaataggaa cttcgctagc ggtaccctgc
     4561 gctctcaggc acgacacgac tcctccgctg cccaccgcag actgaggcag cgctgagtcg
     4621 ccggcgccgc agcgcagatg gtcgcgcccg tgccccccta tctcgcgcct cgcgtggtgc
     4681 ggtccggctg ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag gggagtttgc
     4741 gggggaccgg cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg gcgcgcgccg
     4801 ccaggcgggg gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca cgcgcctcgc
     4861 cgcatcctgt ttcccctccc cctctctgat aggggatgcg caatttgggg aatgggggtt
     4921 gggtgcttgt ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc ccgcctcatc
     4981 ctggtcctaa aacccacttg cactgcgatc gcgggcccgt gcagcatctg cggccgcagg
     5041 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     5101 ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     5161 ctgcaggatt ggatcccaat acgcgtcaat tcactggccg tcgttttaca acgtcgtgac
     5221 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5281 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5341 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5401 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5461 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5521 caagctgtga cagtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5581 cgcgcgagac aaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5641 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5701 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5761 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5821 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5881 aaagatgctg a
//
