LOCUS       sequence-432145-        5707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides NOT intersectional (Flp NOT Cre) 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-CoffFon-TVA(mChr)-W3SL
FEATURES             Location/Qualifiers
     source          1..5707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(103..122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      916..1504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1405..1424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1792..1813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1828..1833,1834..1851,1852..1858)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1866..1882
                     /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     1871..1893
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1890..1906
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1890..1906
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1946..2086
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1957..2086
                     /label=AAV2 ITR
     primer_bind     2459..2479
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2667..2700
                     /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)"
     regulatory      2719..2728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3163..3870
                     /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(3305..3323)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3573..3592
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3891..3924)
                     /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    complement(3975..4008)
                     /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    4101..4134
                     /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    4185..4218
                     /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(4315..4348)
                     /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)"
     promoter        complement(4361..4808)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4842..4971
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     complement(4997..5014)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4997..5013)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5006..5028)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5222..5241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5341..5363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5401..5419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5487..5591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5592..5660,5661..5707,1..745)
                     /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 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
       61 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      121 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      181 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      241 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      301 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      361 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      421 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      481 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      541 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      601 ttctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      661 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      721 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
      781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1681 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1741 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     1801 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1861 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1921 gctcggcgcg ccattgggat ccaatcctgc aggcagctgc gcgctcgctc gctcactgag
     1981 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     2041 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctggag ctccggttac
     2101 cggccgcttt aaaaaacctc ccacatctcc ccctgaacct gaaacataaa atgaatgcaa
     2161 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2221 caaatttcac aaataaagct agatggttac aaataaagca atagcatcac aaatttcaca
     2281 aataaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag
     2341 gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca aggaggagaa
     2401 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     2461 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     2521 cagaggttga ttatctcggg catcgaatat cgaatcggct aagctctaga aagcttcgta
     2581 cggaagttcc tattcttcaa aaggtatagg aacttctagc tggagcaatc tactagctgc
     2641 tgatgtatta caagattgtc tgcttagaag ttcctattct ctagaaagta taggaacttc
     2701 ctcgagaccg gtactagtgc caccatggcg cggctgctgc ccgcgctgct gctgctgctg
     2761 ctgcccggta acgtgaccgg taacgggtcc ggtaacggtt ctttgtcccg ttgccccccc
     2821 ggtcagttcc gctgctcgga gccgcccggt gcccacgggg agtgttaccc gcaggactgg
     2881 ctgtgcgacg gacaccccga ctgcgacgac gggcgggacg agtggggctg cgggaccagc
     2941 gcgatccccg cggtgcccac ggacaacggc acagaggctc ccactgcccc tgctcctgga
     3001 cgtgctctgc cagccaggaa tcacggccgc atgtggatgc tgatcactgc agtgctcctg
     3061 tgctgcctgg tagctgtggg tggtatcgct gcatggggga agtccaaagc aaaaagcagg
     3121 tctgacatct tcagtcttga aagcgcatcc aaggggggtg gggtgagcaa gggcgaggag
     3181 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3241 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3301 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3361 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3421 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3481 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3541 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3601 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3661 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3721 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     3781 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     3841 accggcggca tggacgagct gtacaagtaa gcatgcacat gcatgaattc ataacttcgt
     3901 atagtatacc ttatacgaag ttattctttg cacctatcta aagaataaca gtgataattt
     3961 ctgggttaag gcaaataact tcgtatagga tactttatac gaagttataa aaacaaaaag
     4021 aaaagcgcgt ttcgtcctat ttgggactcg tcagctggat gtacgcgttt ccgcgaatcc
     4081 aggacgcgca aagaaagaaa ataacttcgt ataaggtata ctatacgaag ttattaagcg
     4141 cagctagagg taatacaaca atcttttcaa attgctccag ctacataact tcgtataaag
     4201 tatcctatac gaagttatct taaggtcgac gaagttccta taccttttga agaataggaa
     4261 cttctaagag gtttcatatt gctaatagca gctacaatcc agctaccatt ctgcgaagtt
     4321 cctatacttt ctagagaata ggaacttcgc tagcggtacc ctgcgctctc aggcacgaca
     4381 cgactcctcc gctgcccacc gcagactgag gcagcgctga gtcgccggcg ccgcagcgca
     4441 gatggtcgcg cccgtgcccc cctatctcgc gcctcgcgtg gtgcggtccg gctgggccgg
     4501 cggcggcgcg gacgcgacca aggtggccgg gaaggggagt ttgcggggga ccggcgagtg
     4561 acgtcagcgc gccttcagtg ctgaggcggc ggtggcgcgc gccgccaggc gggggcgaag
     4621 gcactgtccg cggtgctgaa gctggcagtg cgcacgcgcc tcgccgcatc ctgtttcccc
     4681 tccccctctc tgatagggga tgcgcaattt ggggaatggg ggttgggtgc ttgtccagtg
     4741 ggtcggggtc ggtcgtcagg taggcacccc caccccgcct catcctggtc ctaaaaccca
     4801 cttgcactgc gatcgcgggc ccgtgcagca tctgcggccg caggaacccc tagtgatgga
     4861 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4921 ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gattggatcc
     4981 caatacgcgt caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5041 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5101 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     5161 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     5221 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5281 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacagtct
     5341 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacaaaagg
     5401 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5521 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5581 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5701 agttggg
//
