LOCUS       sequence-470110-        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intersectional expression of Myc-tagged synaptophysin in the 
            presence of Cre and Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  TITLE     Arber Lab plasmids March 2025
REFERENCE   2  (bases 1 to 7418)
  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"
COMMENT     Sequence Label: pAAV-Ef1a-Con/Fon-Synaptophysin-10xMyc-WPRE
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(450..483)
                     /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)"
     promoter        complement(490..1668)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          499..1437
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(522..542)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   1769..1898
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1960..2548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2040..2059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2719..3510,3511..3579))
                     /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     3342..3361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3580..3684)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3752..3770
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3808..3830)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3930..3949
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3966..4421)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4103..4124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4315..4334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4496..4636
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4507..4636
                     /label=AAV2 ITR
     polyA_signal    4676..5152
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     4934..4953
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    5184..5772
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5290..5301
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5699..5719
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    5881..5914
                     /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)"
     regulatory      5921..5930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(6157..6190)
                     /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(6241..6274)
                     /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)."
     primer_bind     6387..6408
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    7272..7305
                     /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    7356..7389
                     /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)."
ORIGIN
        1 tgttcagaga acatggttaa ctggttaagt catgtcgtcc cacagctgcc cgtgggacaa
       61 aaactcatct cagaagagga tttagggcag aagctaatat cggaggaaga cttgggtcaa
      121 aaactgatta gtgaagagga tcttggccag aagttaataa gcgaggaaga cctcggacag
      181 aagttgattt ctgaggagga tctagggcaa aaacttatct ccgaagaaga cctgggtcag
      241 aagctaatat cagaggaaga tttaggccaa aaactgattt cggaagagga cttgggacaa
      301 aaattaataa gtgaagaaga tcttgggcag aaattgatta gcgaggagga cctctagggc
      361 gcgccgaagt tcctatacct tttgaagaat aggaacttct aagaggtttc atattgctaa
      421 tagcagctac aatccagcta ccattctgcg aagttcctat actatttgaa gaataggaac
      481 ttcggatcct cacgacacct gaaatggaag aaaaaaactt tgaaccactg tctgaggctt
      541 gagaatgaac caagatccaa actcaaaaag ggcaaattcc aaggagaatt acatcaagtg
      601 ccaagctggc ctaacttcag tctccaccca ctcagtgtgg ggaaactcca tcgcataaaa
      661 cccctccccc caacctaaag acgacgtact ccaaaagctc gagaactaat cgaggtgcct
      721 ggacggcgcc cggtactccg tggagtcaca tgaagcgacg gctgaggacg gaaaggccct
      781 tttcctttgt gtgggtgact cacccgcccg ctctcccgag cgccgcgtcc tccattttga
      841 gctccctgca gcagggccgg gaagcggcca tctttccgct cacgcaactg gtgccgaccg
      901 ggccagcctt gccgcccagg gcggggcgat acacggcggc gcgaggccag gcaccagagc
      961 aggccggcca gcttgagact acccccgtcc gattctcggt ggccgcgctc gcaggccccg
     1021 cctcgccgaa catgtgcgct gggacgcacg ggccccgtcg ccgcccgcgg ccccaaaaac
     1081 cgaaatacca gtgtgcagat cttggcccgc atttacaaga ctatcttgcc agaaaaaaag
     1141 cgtcgcagca ggtcatcaaa aattttaaat ggctagagac ttatcgaaag cagcgagaca
     1201 ggcgcgaagg tgccaccaga ttcgcacgcg gcggccccag cgcccaggcc aggcctcaac
     1261 tcaagcacga ggcgaagggg ctccttaagc gcaaggcctc gaactctccc acccacttcc
     1321 aacccgaagc tcgggatcaa gaatcacgta ctgcagccag gtggaagtaa ttcaaggcac
     1381 gcaagggcca taacccgtaa agaggccagg cccgcgggaa ccacacacgg cacttacctg
     1441 tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata
     1501 cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt
     1561 taccccgcgc caccttctct aggcaccggt tcaattgccg acccctcccc ccaacttctc
     1621 ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagccaa ttcccactcc
     1681 tttcaagacc tagaaggtcc attagctgca aagattcctc tctgtttaaa actttatcca
     1741 tctttgcaaa gcttacgcgt gcggccgcag gaacccctag tgatggagtt ggccactccc
     1801 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     1861 ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa aaggccagca
     1921 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1981 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2041 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2101 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2161 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2221 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2281 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2341 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2401 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2461 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2521 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2581 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2641 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2701 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2761 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2821 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2881 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2941 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3001 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3061 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3121 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3181 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3241 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3301 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3361 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3421 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3481 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3541 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3601 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3661 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3721 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     3781 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     3841 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     3901 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     3961 ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4021 ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     4081 cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4141 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4201 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4261 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4321 gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     4381 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacgt
     4441 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgcccctgc
     4501 aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
     4561 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     4621 atcactaggg gttcctgcgg ccgctcggtc cgcacgtggt tacctacaaa atcagaagga
     4681 cagggaaggg agcagtggtt cacgcctgta atcccagcaa tttgggaggc caaggtgggt
     4741 agatcacctg agattaggag ttggagacca gcctggccaa tatggtgaaa ccccgtctct
     4801 accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc ctggaatccc aacaactcgg
     4861 gaggctgagg caggagaatc gcttgaaccc aggaggcgga gattgcagtg agccaagatt
     4921 gtgccactgc actccagctt ggttcccaat agaccccgca ggccctacag gttgtcttcc
     4981 caacttgccc cttgctccat accacccccc tccaccccat aatattatag aaggacacct
     5041 agtcagacaa aatgatgcaa cttaatttta ttaggacaag gctggtgggc actggagtgg
     5101 caacttccag ggccaggaga ggcactgggg aggggtcaca gggatgccac ccgtagatct
     5161 ctcgagcagc gctcggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
     5221 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
     5281 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
     5341 ggcaacatag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
     5401 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     5461 cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc
     5521 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
     5581 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
     5641 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     5701 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     5761 cagaggttga ttatcgataa gcttgatatc gaattcgaag ttcctattct tcaaaaggta
     5821 taggaacttc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
     5881 gaagttccta ttcttcaaat agtataggaa cttctctaga gccaccatgg acgtggtgaa
     5941 tcagctggtg gctgggggtc agttccgggt ggtcaaggag ccccttggct tcgtgaaggt
     6001 gctgcagtgg gtctttgcca tcttcgcctt tgctacgtgt ggcagctaca ccggggagct
     6061 tcggctgagc gtggagtgtg ccaacaagac ggagagtgcc ctcaacatcg aagttgaatt
     6121 cgagtacccc ttcaggtacg tgtcggggtt gctagcataa cttcgtatag tataccttat
     6181 acgaagttat tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
     6241 ataacttcgt ataggatact ttatacgaag ttatggtacc cctcctgtac tcaccttgcc
     6301 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     6361 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     6421 cccggtgaac agctcctcgc ccttgctcac ggcggccgcc tgattggaga aggaggtggg
     6481 cgcaccctgt tggccatagc cttgctgccc atagtcgccc tgaggcccgt agccaccgcc
     6541 accgctggct ggctgcccgt aatcgggttg ataaccaccc tgaggcccgt aggagtcctg
     6601 gggcccatag cctccggggc cctgcccgta gcccgcatcg ccgtaggcat cgccaggtgc
     6661 tggttgcttt tccggggcgc ctggaggtgc gcgcatgaat ggggctgccc agcctgtctc
     6721 cttgaacacg aaccataagt tgccaaccca gagcaccagg ttcaggaagc caaacaccac
     6781 tgtggtgttg agtcctgaag tcacagggtc cctcagttcc ttgcatgtgt tccctgtctg
     6841 gcggcacatg ggcatctcct tgataatgtt ctctgggtcc gtggccatct tcacatcgga
     6901 caggcctttg gcccaggctg atgaactaac tagccacatg aaagcgaaca cggctgtagc
     6961 cagaaagtcc atcataggcc ctttgttgtt ctctcggtac ttgttctgca ggaagatgta
     7021 ggtggccagg gcccccatgg agtagaggaa ggcaaacaca gccacggtga caaagaattc
     7081 agccgacgag gagtagtccc caaccaggaa gatcttggta gtgccccctt tgacgcagga
     7141 gggtgcatca aagtacactt ggtgcagcct aaacacatga gaaatagagt cataagcaca
     7201 tgggtcacat acaggagata tgtatataac attaatacaa ttttataaaa aaaagggggg
     7261 gcacaactag tataacttcg tataaggtat actatacgaa gttattaaga ggtttcatat
     7321 tgctaatagc agctacaatc cagctaccat tctgcataac ttcgtataaa gtatcctata
     7381 cgaagttatg tcgactggag agtggccagc ccttctca
//
