LOCUS       sequence-432146-        6221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides AND intersectional (Cre+Flp) expression of HA-hM3Dq.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6221)
  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 6221)
  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)hM3Dq-W3SL
FEATURES             Location/Qualifiers
     source          1..6221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(379..483)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(607..629)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     729..748
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     942..964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     956..973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     957..973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   999..1128
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1162..1609
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    1718..1765
                     /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)."
     protein_bind    complement(1778..1811)
                     /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(1904..1937)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3709..3735)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3854..3901)
                     /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)."
     primer_bind     complement(4005..4025)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4398..4538
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4398..4527
                     /label=AAV2 ITR
     primer_bind     complement(4578..4594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4578..4594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4591..4613)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4602..4618
                     /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)."
     promoter        complement(join(4626..4632,4633..4650,4651..4656))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4671..4692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4980..5568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5060..5079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5739..6221,1..309,310..378))
                     /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 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
       61 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      121 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      181 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      241 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      301 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      361 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      421 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      481 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      541 aacctataaa aataggcgta tcacgaggcc cttttgtctc gcgcgtttcg gtgatgacgg
      601 tgaaaacctc tgacacatgc agctcccgga gactgtcaca gcttgtctgt aagcggatgc
      661 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      721 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
      781 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
      841 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      901 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      961 aacgacggcc agtgaattga cgcgtattgg gatccaatcc tgcaggcagc tgcgcgctcg
     1021 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     1081 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcagat
     1141 gctgcacggg cccgcgatcg cagtgcaagt gggttttagg accaggatga ggcggggtgg
     1201 gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca
     1261 aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc
     1321 actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg
     1381 ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc
     1441 cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg
     1501 cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc
     1561 tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagg gtaccgctag
     1621 cgaagttcct attcttcaaa aggtatagga acttcatcaa aataggaaga ccaatgcttc
     1681 accatcgacc cgaattgcca agcatcacca tcgacacgaa gttcctattc cgaagttcct
     1741 attctctaga aagtatagga acttcgtcga ccttaagata acttcgtata gcatacatta
     1801 tacgaagtta taaaaacaaa aagaaaagcg cgtttcgtcc tatttgggac tcgtcagctg
     1861 gatgtacgcg tttccgcgaa tccaggacgc gcaaagaaag aaaataactt cgtatagcat
     1921 acattatacg aagttatgct acaaggcctg ctcgggtgcg cgcttgtgaa aaatgaccga
     1981 ctgtctctgc tggtactgct gcttgcgcct cttttttttg tcacactggc acagcagcag
     2041 catcttgaaa gtggttctga atgttttgtt gcacagagca tagcacacgg ggttcacggt
     2101 gctgttgatg tagcacagcc agtagcccag attccaaaag gttttgggta tgcagctgtc
     2161 acaaaaggtg ttcaccagaa ccatgatgtt gtatggggtc caagtgatga tgaaggcaag
     2221 caagatcgca ctgagggtct gggccgcttt cttctccttg accagggaca tccttttccg
     2281 cttagtgatc tgacttctgg tcttcagagc aaacctcttg gccagagtgg cttccttgaa
     2341 ggacagaggt agagtggccg tgctcttacc cactgaggag ttgacgtcag aagtcttagc
     2401 tgtgtccacg gctgactcta gctggatggg aagcttggag aagctttttg gaaaactgcc
     2461 tccatcgtcc acgctcttct gggcctgcag cttgtcggct ttcctctcca agtccaccat
     2521 ccccagctcc tcctcaggca cctgcaggtt gtccgatgag ggtaacttgg tggagttgag
     2581 gatggtgctg tgacccggaa gcttgagcac gatggagtag atggctctcg tctcggagcc
     2641 aatgtcctcc tcgtcggagg aggcggagtt ctccagggag gcagcagcat cattgttgtt
     2701 ccaactgtca ctgctgctgt ggtcttggtc catctgctcg gagctgggtt tccagctctt
     2761 ggttgtgaac cagaagtggc agcggccata cttcctcctg ttggagcgtt tcatgctttg
     2821 ctgttgaagt tcgtaactgc tgcagcttcg agaactgccc gtggggtgga caaagttttc
     2881 tgtctctgcc tctgtcccag aggcttgcag gccagcaagc tctttggtac gcttttcagt
     2941 ttccttatag atcctccagt ataaaatagt cataatggtg acaggcatat aaaaaccagc
     3001 gatggctgtg ccaaaagtaa tggtgggctc actgaggaac tgaatgaagc actctcccgg
     3061 aggcacagtt ctctttccaa caaagtattg ccagaacaag atggcaggag cccaaaggac
     3121 aaaggagatg acccaagcca gaccgatcat cacaccggct ctctttgttg ttcgtttggc
     3181 tcggtacgtg agcggcctcg tgatggaaaa gtatctgtca aagctgatga ccagaagatt
     3241 cataacagag gcattgctgg ctacgcagtc aatggcaagc cagaggtcac aggccaagtt
     3301 ccctaaggcc catcgattca tgatgatgta ggtcgtaaac agattcattg aaatgacccc
     3361 gataatcaga tcggcacagg ccaggcttaa gaggaagtag ttgttgaccg tcttcagctg
     3421 cttgttgacc ttaaatgaca caattaccag gatgttgccg atgatggtca ccaaggccag
     3481 gatgcccgtt aagaaagcga tgaagaccac ttgccagacg gtatgacctc ccagagggtc
     3541 atcggtggta ccgtctggag aggagaaatt gccagctgct cgagaaacat tgtagctgcc
     3601 gaaatgagtg acggttcccg ggggcagccc tgcatcggag gggctgtgta tccaggagga
     3661 gctgatgttt ggaaacaaag gcgaggttgt actgttattg tgcaaggtag cgtaatctgg
     3721 aacatcgtat gggtacatgg tggctaccgg tctcgaggaa gttcctatac cttttgaaga
     3781 ataggaactt catcaaaata ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca
     3841 tcaccatcga cacgaagttc ctatactttc tagagaatag gaacttcgga atagtaactt
     3901 ccgtacgaag ctttctagag cttagccgat tcgatattcg atgcccgaga taatcaacct
     3961 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4021 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4081 attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact catcgccgcc
     4141 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttt
     4201 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca tctagcttta tttgtgaaat
     4261 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4321 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcgg
     4381 ccggtaaccg gagctccagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4441 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4501 ggcctcagtg agcgagcgag cgcgcagctg cctgcaggat tggatcccaa tggcgcgccg
     4561 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4621 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4681 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4741 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4801 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4861 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4921 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4981 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5041 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5101 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5161 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5221 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5281 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5341 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5401 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5461 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5521 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5581 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5641 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5701 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5761 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5821 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5881 gaaccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5941 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6001 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6061 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6121 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6181 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca t
//
