LOCUS       sequence-198809-        5843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-on/Flp-on EYFP under the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 5843)
  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"; doi: 
            "10.1038/nmeth.2996"; journalName: "Nat Methods"; date: "2014-07"; 
            volume: "11"; issue: "7"; pages: "763-72"
COMMENT     Sequence Label: pAAV-hSyn Con/Fon EYFP
FEATURES             Location/Qualifiers
     source          1..5843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(249..282)
                     /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(333..366)
                     /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     479..500
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      542..551
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    556..589
                     /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    640..673
                     /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    680..713
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     CDS             complement(809..1060)
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     complement(851..872)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1200..1219
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1441..1474)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    1583..2171
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1636..1656)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2054..2065)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2203..2679
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2402..2421)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2719..2859
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2719..2848
                     /label=AAV2 ITR
     rep_origin      2934..3389
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3021..3040)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3231..3252
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3406..3425)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3525..3547
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3585..3603)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3671..3775
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3776..3844,3845..4636)
                     /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(3994..4013)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4807..5395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5296..5315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5457..5586
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(5631..5843,1..235)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc
       61 ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc
      121 gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc
      181 gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgag
      241 aaggatccat aacttcgtat aggatacttt atacgaagtt atgtagctgg agcaatttga
      301 aaagattgtt gtattacctc tagctgcgct taataacttc gtatagtata ccttatacga
      361 agttatacta gtcctcctgt actcaccttg ccggtggtgc agatgaactt cagggtcagc
      421 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
      481 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
      541 accatggtgg ctagcataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc
      601 cagaaattat cactgttatt ctttagatag gtgcaaagaa taacttcgta taaggtatac
      661 tatacgaagt tatggtaccg aagttcctat tcttcaaata gtataggaac ttcaattgtt
      721 attggtgcaa agaatcactt ccttaagtct ttatactgcc caggaagttc ctattcttca
      781 aaaggtatag gaacttcggc gcgccttact tgtacagctc gtccatgccg agagtgatcc
      841 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
      901 tcagggcgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
      961 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     1021 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     1081 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1141 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1201 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1261 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
     1321 ggccgtagcc gaaggtggtc acgagggtgg gccagggcac gggcagctgt gggacgacat
     1381 gacttaacca gttaaccatg ttctctgaac atgagaaggg ctggccactc tccagagctc
     1441 gaagttccta tactatttga agaataggaa cttcttgcct taacccagaa attatcactg
     1501 ttattcttta gaatggtgca aagagaagtt cctatacctt ttgaagaata ggaacttcga
     1561 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     1621 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1681 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1741 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     1801 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     1861 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     1921 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     1981 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     2041 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2101 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2161 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     2221 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2281 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2341 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2401 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     2461 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     2521 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     2581 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     2641 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     2701 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2761 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2821 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2881 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2941 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3001 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3061 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3121 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3181 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3241 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3301 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3361 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3421 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3481 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3541 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3601 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3721 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3781 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3841 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3901 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3961 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4021 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4081 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4141 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4201 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4261 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4321 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4381 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4441 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4501 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4561 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4621 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4681 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4801 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4861 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5281 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5341 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5401 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg
     5461 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc
     5521 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     5581 gttcctgcgg ccgcacgcgt gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg
     5641 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca
     5701 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
     5761 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
     5821 tcgcccccgc ctggcggcgc gcg
//
