LOCUS       sequence-432136-        6282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides AND intersectional (Cre+Flp) expression of 
            stGtACR1-FusionRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6282)
  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 6282)
  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-stGtACR1(FusionRed)-W3SL
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..153
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   24..153
                     /label=AAV2 ITR
     primer_bind     526..546
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    650..697
                     /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             1772..2467
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMLMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     protein_bind    2675..2708
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2801..2834
                     /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(2847..2894)
                     /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(3003..3450)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3484..3613
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     complement(3639..3656)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3639..3655)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3648..3670)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3864..3883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3983..4005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4043..4061)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4129..4233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4234..4302,4303..5094)
                     /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     complement(4452..4471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5265..5853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5754..5773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6141..6162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6177..6182,6183..6200,6201..6207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6215..6231
                     /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     6220..6242
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6239..6255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6239..6255
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttgggatcca atcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
       61 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
      121 agggagtggc caactccatc actaggggtt cctggagctc cggttaccgg ccgctttaaa
      181 aaacctccca catctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
      241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
      301 taaagctaga tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaacaccacg
      361 gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg
      421 agttccgccg tggcaagaac taaccaggat ttatacaagg aggagaaaat gaaagccata
      481 cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga
      541 gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta
      601 tctcgggcat cgaatatcga atcggctaag ctctagaaag cttcgtacgg aagttactat
      661 tccgaagttc ctattctcta gaaagtatag gaacttcgtg tcgatggtga tgcttggcaa
      721 ttcgggtcga tggtgaagca ttggtcttcc tattttgatg aagttcctat tcttcaaaag
      781 gtataggaac ttcctcgaga ccggtactag tgccaccatg agcagcatta cttgtgaccc
      841 agccatctat ggagagtgga gcagagagaa ccagttttgc gtcgaaaaga gcctgattac
      901 cctggacggc atcaagtacg tgcagctggt catggccgtg gtgagcgcct gccaggtgtt
      961 ctttatggtc actagggcac ccaaagtgcc ttgggaggcc atctacctgc caaccacaga
     1021 aatgatcacc tattcactgg ctttcacagg caacggatac attcgagtgg caaatgggaa
     1081 gtatctgcct tgggctagaa tggcaagttg gctgtgcaca tgtccaatca tgctgggcct
     1141 ggtgagcaac atggccctgg tgaagtacaa atccattcca ctgaatccca tgatgattgc
     1201 cgccagcagc atctgcacag tgttcggaat cactgcctcc gtggtcctgg accccctgca
     1261 cgtgtggctg tactgtttca tctctagtat tttctttatc tttgagatgg tggtcgcatt
     1321 cgccattttt gccatcacta ttcatgattt tcagaccatc ggaagtccca tgtcactgaa
     1381 ggtggtcgaa aggctgaaac tgatgagaat cgtgttctac gtgagctgga tggcctatcc
     1441 tatcctgtgg agtttttcaa gcacaggggc ttgcattatg tctgagaaca cttcctctgt
     1501 gctgtacctg ctgggcgacg cactgtgcaa aaatacctat ggaatcctgc tgtgggctac
     1561 tacctgggga ctgctgaacg gcaagtggga ccgagattac gtgaaaggac gcaatgtcga
     1621 tgggacactg atgcccgaat atgaacagga tctggagaaa ggcaatactg aacgatatga
     1681 ggatgctcgg gctggggaaa ccgccgccaa gagcaggatc accagcgagg gcgagtacat
     1741 ccccctggac cagatcgaca tcaacgtggt gatggtgagc gagctgatta aggagaacat
     1801 gctcatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga
     1861 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtcg tcgagggcgg
     1921 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt
     1981 catcaagcac cctccgggca tccccgactt ctttaagcag tccttccctg agggcttcac
     2041 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
     2101 cctccaggac ggctgcctca tctacaacgt caaggttaga ggggtgaact tcccagccaa
     2161 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga cgatgtaccc
     2221 tgctgacgga ggactggaag gcgcatgtga catggctctg aagctcgtgg gaggaggaca
     2281 cctgatctgc aaccttgaga caacatacag atccaagaaa cccgctacga acctcaagat
     2341 gcccggcgtc tacaacgtgg accacagact ggaaagaatc aaggaggccg acgatgagac
     2401 atacgtcgag cagcacgagg tggctgtggc cagatactct actggtggcg ctggtgatgg
     2461 aggtaaacag tcccagccta ttctgaacac caaagagatg gccccccaga gtaaaccccc
     2521 tgaggagctg gaaatgtcct caatgccctc ccctgtggca cctcttcctg caagaacaga
     2581 gggcgtgatc gacatgaggt ccatgagttc tattgactct tttatctcat gcgccaccga
     2641 ttttccagag gctacgcggt ttaagtaaga attcataact tcgtataatg tatgctatac
     2701 gaagttattt tctttctttg cgcgtcctgg attcgcggaa acgcgtacat ccagctgacg
     2761 agtcccaaat aggacgaaac gcgcttttct ttttgttttt ataacttcgt ataatgtatg
     2821 ctatacgaag ttatcttaag gtcgacgaag ttcctatact ttctagagaa taggaacttc
     2881 ggaataggaa cttcgtgtcg atggtgatgc ttggcaattc gggtcgatgg tgaagcattg
     2941 gtcttcctat tttgatgaag ttcctatacc ttttgaagaa taggaacttc gctagcggta
     3001 ccctgcgctc tcaggcacga cacgactcct ccgctgccca ccgcagactg aggcagcgct
     3061 gagtcgccgg cgccgcagcg cagatggtcg cgcccgtgcc cccctatctc gcgcctcgcg
     3121 tggtgcggtc cggctgggcc ggcggcggcg cggacgcgac caaggtggcc gggaagggga
     3181 gtttgcgggg gaccggcgag tgacgtcagc gcgccttcag tgctgaggcg gcggtggcgc
     3241 gcgccgccag gcgggggcga aggcactgtc cgcggtgctg aagctggcag tgcgcacgcg
     3301 cctcgccgca tcctgtttcc cctccccctc tctgataggg gatgcgcaat ttggggaatg
     3361 ggggttgggt gcttgtccag tgggtcgggg tcggtcgtca ggtaggcacc cccaccccgc
     3421 ctcatcctgg tcctaaaacc cacttgcact gcgatcgcgg gcccgtgcag catctgcggc
     3481 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3541 aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc
     3601 agctgcctgc aggattggat cccaatacgc gtcaattcac tggccgtcgt tttacaacgt
     3661 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3721 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3781 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3841 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3901 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3961 tacagacaag ctgtgacagt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4021 ccgaaacgcg cgagacaaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4081 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4201 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4261 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4321 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4381 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4441 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4501 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4561 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4621 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4681 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4741 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4801 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4861 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4921 gctgataaat ctggagccgg tgagcgtggt tctcgcggta tcattgcagc actggggcca
     4981 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5041 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5101 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5941 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6001 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6061 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6121 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6181 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6241 ggaaacagct atgaccatga ttacgccaag ctcggcgcgc ca
//
