LOCUS       sequence-350686-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Viral vector for co-expression of a CDS of interest and mCherry in 
            cells expressing Flp AND NOT Cre driven by a Synapsin promoter. 
            Contains an in-frame P2A sequence and an MCS for cloning of the 
            CDS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  AUTHORS   Ubina T, Vahedi-Hunter T, Agnew-Svoboda W, Wong W, Gupta A, 
            Santhakumar V, Riccomagno MM
  TITLE     ExBoX - a simple Boolean exclusion strategy to drive expression in 
            neurons.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272538. doi: 
            10.1242/jcs.257212. Epub 2021 Oct 20.
  PUBMED    34515305
REFERENCE   2  (bases 1 to 5520)
  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: "J Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272538. doi"; 
            pages: " 10.1242/jcs.257212. Epub 2021 Oct 20"
COMMENT     Sequence Label: pAAV-SynI-FlpOn-CreOff-MCS-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..5520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..191
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   51..180
                     /label=AAV2 ITR
     rep_origin      266..721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(353..372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     563..584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(738..757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     857..879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(917..935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1003..1107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1108..1176,1177..1968)
                     /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(1326..1345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2139..2727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2628..2647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2789..2929
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2800..2929
                     /label=AAV2 ITR
     protein_bind    2936..2956
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2978..3425
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    3425..3449
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    3540..3573
                     /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    3574..3607
                     /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)."
     CDS             complement(3616..4326)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3894..3913)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4163..4181
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4327..4383)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     protein_bind    4544..4577
                     /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(4662..4695)
                     /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(4698..4722)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4732..5320
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4785..4805)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5203..5214)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5350..5367)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(5356..5520,1..43)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagggccgc aggaacccct
       61 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
      121 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
      181 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
      241 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
      301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
      361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      481 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
      601 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
      661 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
      721 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
      781 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
      841 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
      901 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
      961 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1021 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1081 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1141 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1201 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1261 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1321 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1381 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1441 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1501 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1561 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1621 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1681 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1741 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1801 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     1861 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1921 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1981 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2041 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2101 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2161 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2221 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2281 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2341 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2401 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2461 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2521 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2581 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2641 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2701 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2761 gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact
     2821 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
     2881 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt ctagacaact
     2941 ttgtatagaa aagttgctgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
     3001 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
     3061 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
     3121 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
     3181 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
     3241 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
     3301 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
     3361 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
     3421 cgcagcaagt ttgtacaaaa aagcaggctg ccaccgaagt tcctattcta tcagaagtat
     3481 aggaacttcg cagaatggta gctggattgt agctgctatt agcaatatga aacctcttag
     3541 aagttcctat tctctagaaa gtataggaac ttcataactt cgtataaagt atcctatacg
     3601 aagttattta attaattact tgtacagctc gtccatgccg ccggtggagt ggcggccctc
     3661 ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt
     3721 gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt
     3781 gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt
     3841 cagggcgccg tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt
     3901 ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa
     3961 ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
     4021 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc
     4081 ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca
     4141 ggcgaagggc agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta
     4201 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
     4261 gtgcaccttg aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct
     4321 caccataggc ccggggtttt cttcaacatc tcctgcttgc tttaacagag agaagttcgt
     4381 ggctccgctt ccactagtac tcgagtcccc gcggccggcc tgcttcactt aattaacaat
     4441 ggtgggagat ctgcagtggt gggtaacctg ttctcccggg atgaaggatc cgcgaagctt
     4501 gggtcgacgg tacccctcta gagctagcta cgtaggcgcg ccaataactt cgtataaagt
     4561 atcctatacg aagttatgaa gttcctatac ttctgataga ataggaactt cttgccttaa
     4621 cccagaaatt atcactgtta ttctttagaa tggtgcaaag agaagttcct atactttcta
     4681 gagaatagga acttccaacc cagctttctt gtacaaagtg ggaattccga taatcaacct
     4741 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4801 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4861 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4921 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4981 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5041 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5101 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     5161 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5221 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5281 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgggaatt cctagagctc
     5341 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5401 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5461 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
//
