LOCUS       sequence-432150-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides NOT intersectional (Cre NOT Flp) expression of TVA-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5707)
  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 5707)
  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-ConFoff-TVA(mChr)-W3SL
FEATURES             Location/Qualifiers
     source          1..5707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(259..363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     431..449
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(487..509)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     609..628
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     822..844
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     836..853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     837..853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   879..1008
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        1042..1489
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(1502..1535)
                     /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(1586..1619)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    1632..1665
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(1842..1875)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1980..2687)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2258..2277)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2527..2545
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3122..3131
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3150..3183
                     /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    3234..3267
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     primer_bind     complement(3371..3391)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3764..3904
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3764..3893
                     /label=AAV2 ITR
     primer_bind     complement(3944..3960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3944..3960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3957..3979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3968..3984
                     /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(3992..3998,3999..4016,4017..4022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4037..4058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4346..4934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4426..4445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5105..5707,1..189,190..258))
                     /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 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
       61 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      361 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      421 aacctataaa aataggcgta tcacgaggcc cttttgtctc gcgcgtttcg gtgatgacgg
      481 tgaaaacctc tgacacatgc agctcccgga gactgtcaca gcttgtctgt aagcggatgc
      541 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      601 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
      661 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
      721 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      781 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      841 aacgacggcc agtgaattga cgcgtattgg gatccaatcc tgcaggcagc tgcgcgctcg
      901 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      961 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcagat
     1021 gctgcacggg cccgcgatcg cagtgcaagt gggttttagg accaggatga ggcggggtgg
     1081 gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca
     1141 aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc
     1201 actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg
     1261 ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc
     1321 cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg
     1381 cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc
     1441 tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagg gtaccgctag
     1501 cataacttcg tataggatac tttatacgaa gttatgtagc tggagcaatt tgaaaagatt
     1561 gttgtattac ctctagctgc gcttaataac ttcgtatagt ataccttata cgaagttatg
     1621 tcgaccttaa ggaagttcct attctctaga aagtatagga acttcgcaga atggtagctg
     1681 gattgtagct gctattagca atatgaaacc tcttagaagt tcctattctt caaaaggtat
     1741 aggaacttct ttctttcttt gcgcgtcctg gattcgcgga aacgcgtaca tccagctgac
     1801 gagtcccaaa taggacgaaa cgcgcttttc tttttgtttt tgaagttcct atactttcta
     1861 gagaatagga acttctaagc agacaatctt gtaatacatc agcagctagt agattgctcc
     1921 agctagaagt tcctatacct tttgaagaat aggaacttcg aattcatgca tgtgcatgct
     1981 tacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt
     2041 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag
     2101 gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag
     2161 tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg
     2221 gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg
     2281 tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg
     2341 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac
     2401 ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg
     2461 taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg
     2521 ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
     2581 tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc
     2641 atgaactcct tgatgatggc catgttatcc tcctcgccct tgctcacccc accccccttg
     2701 gatgcgcttt caagactgaa gatgtcagac ctgctttttg ctttggactt cccccatgca
     2761 gcgataccac ccacagctac caggcagcac aggagcactg cagtgatcag catccacatg
     2821 cggccgtgat tcctggctgg cagagcacgt ccaggagcag gggcagtggg agcctctgtg
     2881 ccgttgtccg tgggcaccgc ggggatcgcg ctggtcccgc agccccactc gtcccgcccg
     2941 tcgtcgcagt cggggtgtcc gtcgcacagc cagtcctgcg ggtaacactc cccgtgggca
     3001 ccgggcggct ccgagcagcg gaactgaccg ggggggcaac gggacaaaga accgttaccg
     3061 gacccgttac cggtcacgtt accgggcagc agcagcagca gcagcgcggg cagcagccgc
     3121 gccatggtgg cactagtacc ggtctcgaga taacttcgta taaagtatcc tatacgaagt
     3181 tatttgcctt aacccagaaa ttatcactgt tattctttag ataggtgcaa agaataactt
     3241 cgtataaggt atactatacg aagttatcgt acgaagcttt ctagagctta gccgattcga
     3301 tattcgatgc ccgagataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3361 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3421 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttagttc
     3481 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3541 tgggcactga caattccgtg gtgtttattt gtgaaatttg tgatgctatt gctttatttg
     3601 taaccatcta gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3661 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3721 gagatgtggg aggtttttta aagcggccgg taaccggagc tccaggaacc cctagtgatg
     3781 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3841 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     3901 caggattgga tcccaatggc gcgccgagct tggcgtaatc atggtcatag ctgtttcctg
     3961 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4021 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4081 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5221 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa
     5281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5401 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5701 gttgctc
//
