LOCUS       sequence-466934-        5700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides AND intersectional (Cre+Flp) expression of GCaMP8m.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5700)
  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 5700)
  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-GC8m-W3SL
FEATURES             Location/Qualifiers
     source          1..5700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      917..1505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1406..1425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1793..1814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1829..1834,1835..1852,1853..1859)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1867..1883
                     /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     1872..1894
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1891..1907
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1891..1907
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1947..2087
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1958..2087
                     /label=AAV2 ITR
     primer_bind     2460..2480
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2584..2631
                     /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             2755..2772
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     3054..3075
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3181..3202)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3442..3461)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    4027..4060
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4153..4186
                     /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(4199..4246)
                     /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(4355..4802)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4836..4965
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     complement(4991..5008)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4991..5007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5000..5022)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5216..5235)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5335..5357
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5395..5413)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5481..5585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5586..5654,5655..5700,1..746)
                     /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 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
       61 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      601 gttctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
      781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1561 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1621 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1681 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1741 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     1801 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1861 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1921 agctcggcgc gccattggga tccaatcctg caggcagctg cgcgctcgct cgctcactga
     1981 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     2041 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgga gctccggtta
     2101 ccggccgctt taaaaaacct cccacatctc cccctgaacc tgaaacataa aatgaatgca
     2161 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2221 acaaatttca caaataaagc tagatggtta caaataaagc aatagcatca caaatttcac
     2281 aaataaacac cacggaattg tcagtgccca acagccgagc ccctgtccag cagcgggcaa
     2341 ggcaggcggc gatgagttcc gccgtggcaa gaactaacca ggatttatac aaggaggaga
     2401 aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac
     2461 atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat
     2521 ccagaggttg attatctcgg gcatcgaata tcgaatcggc taagctctag aaagcttcgt
     2581 acggaagtta ctattccgaa gttcctattc tctagaaagt ataggaactt cgtgtcgatg
     2641 gtgatgcttg gcaattcggg tcgatggtga agcattggtc ttcctatttt gatgaagttc
     2701 ctattcttca aaaggtatag gaacttcctc gagaccggta ctagtgccac catgcatcat
     2761 caccatcatc acacgcgtcg caagaagacc ttcaaggagg tggccaccgc cgtgaagatc
     2821 atcggccgcc tgatgggcct gaagatcaac gtctatatca aggccgacaa gcagaagaac
     2881 ggcatcaagg cgaacttcca catccgccac aacatcgagg acggcggcgt gcagctcgcc
     2941 taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3001 tacctgagcg tggagtccaa actttcgaaa gaccccaacg agaagcgcga tcacatggtc
     3061 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     3121 ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3181 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggtgag
     3241 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3301 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3361 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacatccag
     3421 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3481 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3541 aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga agagcagatc
     3601 gcagaataca aagaggcttt ctccctattt gacaaggacg gggatgggac aataacaacc
     3661 aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc agagctgcag
     3721 gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc tgagttcctg
     3781 acaatgatgg caagaaaaat gaaatacagg gacacggaag aagaaattag agaagcgttc
     3841 ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg ccacgtgatg
     3901 acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag ggaagcagac
     3961 atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac agcgaagtga
     4021 gaattcataa cttcgtataa tgtatgctat acgaagttat tttctttctt tgcgcgtcct
     4081 ggattcgcgg aaacgcgtac atccagctga cgagtcccaa ataggacgaa acgcgctttt
     4141 ctttttgttt ttataacttc gtataatgta tgctatacga agttatctta aggtcgacga
     4201 agttcctata ctttctagag aataggaact tcggaatagg aacttcgtgt cgatggtgat
     4261 gcttggcaat tcgggtcgat ggtgaagcat tggtcttcct attttgatga agttcctata
     4321 ccttttgaag aataggaact tcgctagcgg taccctgcgc tctcaggcac gacacgactc
     4381 ctccgctgcc caccgcagac tgaggcagcg ctgagtcgcc ggcgccgcag cgcagatggt
     4441 cgcgcccgtg cccccctatc tcgcgcctcg cgtggtgcgg tccggctggg ccggcggcgg
     4501 cgcggacgcg accaaggtgg ccgggaaggg gagtttgcgg gggaccggcg agtgacgtca
     4561 gcgcgccttc agtgctgagg cggcggtggc gcgcgccgcc aggcgggggc gaaggcactg
     4621 tccgcggtgc tgaagctggc agtgcgcacg cgcctcgccg catcctgttt cccctccccc
     4681 tctctgatag gggatgcgca atttggggaa tgggggttgg gtgcttgtcc agtgggtcgg
     4741 ggtcggtcgt caggtaggca cccccacccc gcctcatcct ggtcctaaaa cccacttgca
     4801 ctgcgatcgc gggcccgtgc agcatctgcg gccgcaggaa cccctagtga tggagttggc
     4861 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4921 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggattgg atcccaatac
     4981 gcgtcaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5041 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5101 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     5161 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     5221 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5281 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgaca gtctccggga
     5341 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacaa aagggcctcg
     5401 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
//
