LOCUS       Exported                6922 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  This AAV plasmid in the presence of Cre recombinase expresses an 
            extracellular SYFP2 fused to the neuroligin-1 cytoplasmic domain for
            targeting to the synapse with a cell fill dTomato..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.hSyn.FLEX.SYFP2-POST-T2A-dTomato.FLEX.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6922)
  AUTHORS   Kuljis DA, Park E, Telmer CA, Lee J, Ackerman DS, Bruchez MP, Barth 
            AL
  TITLE     Fluorescence-based quantitative synapse analysis for cell-type 
            specific connectomics.
  JOURNAL   eNeuro. 2019 Sep 23. pii: ENEURO.0193-19.2019. doi: 
            10.1523/ENEURO.0193-19.2019.
  PUBMED    31548370
REFERENCE   2  (bases 1 to 6922)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6922
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   2..131
                     /label=AAV2 ITR
     promoter        213..660
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(690..723)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(786..819)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1583..1636)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2033..2044
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(3071..3100)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     sig_peptide     complement(3107..3169)
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     protein_bind    3204..3237
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    3308..3341
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3383..3971
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3436..3456)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3854..3865)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3974..3990)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3975..3991)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4020..4141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4031..4050)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4085..4104
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4476..4493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4647..5235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4727..4746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5406..6266)
                     /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     6029..6048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6267..6371)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6397..6852)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6534..6555)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6746..6765
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      181 taggaagatc tctgcagagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg
      241 aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc
      301 cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga
      361 ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc
      421 gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa
      481 actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac
      541 cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac
      601 tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag
      661 tcgaattcaa gctgctagca aggatcccca taacttcgta taaagtatcc tatacgaagt
      721 tatatcaaaa taggaagacc aatgcttcac catcgacccg aattgccaag catcaccatc
      781 gacccataac ttcgtataat gtatgctata cgaagttata ctagctagag atatcgtcga
      841 cggccgcttt acttgtacat tattacttat acagctcatc catgccatac aagaacaggt
      901 gatgccgacc ttctgagcgt tcgtactgct cgacgattgt gtagtcctcg ttgtgactcg
      961 tgatgtccaa cttggtgtcc acgtagtagt agccaggcag ctgaaccggc tttttggcca
     1021 tatatatagt tttgaactca accaggtagt gcccgccatc tttgagttta agcgcctgat
     1081 ggatttcgcc tttaagaacg ccatcgcggg ggtacagtcg ttctgtagag gcctcccacc
     1141 ccatggtctt cttctgcatc acggggccat cggggggaaa atttgttccg cgcatcttca
     1201 ccttgtaaat cagggtgccg tcctgcagag aggagtcttg cgtgacagtg accagcccac
     1261 catcctcaaa attcattact cgttcccact taaatccttc aggaaatgac agttttttgt
     1321 aatcaggtat atcagcaggg tgtttgacgt aagccttgct accgtacatg aattgagggc
     1381 tgaggatgtc ccaggcgaaa ggaagggggc cacctttggt aaccttgagc ttggcggttt
     1441 gtgtgccctc atatggtctt ccttcaccct caccttcaat ttcaaactcg tgcccgttca
     1501 tgctaccctc cattctaact ttgaatctca taaactcctt aattacttcc tcacccttag
     1561 acaccattgt cgaccccggg gccggtccgg gattctcctc gacgtcgcca catgtgagca
     1621 gagagccccg accctcagat ctgccaccgc tgccacccac ccgtgttgtg ctgtgggagt
     1681 gggggtgtgg atgcggatga gggagagtat tattctgtcc accagtgaat gtgttgaaag
     1741 tgtggagagg ctggattcct ggaatagtgt tgggaatcat ggttatggta ttgggtgtca
     1801 tcaggggaat gtcgtctggg cttcggcgca ttgccagggt atagtccgga gggcatgctg
     1861 tcctcagcac cacctcatgg ggatggatgg attcacactc gtggtccagg tctgtgtgtt
     1921 tcatctgaag actcatgatc tcctcctcag gtgcgtgcgt aagatcattg gttgtagttc
     1981 gttgcggact acatctccta tgtacgtcgt ggcgcctttt gtccttcttg taatagaggg
     2041 ccgcgaaagc cagaatgttc aggaaaagca gggaagctcc tacagctatg gtcacggaca
     2101 gctctgtgct gtagtccctt tggtctacgc taaatggact aggctgttgt ttggggtcgt
     2161 cttgcttagc ggttggaaat gcgctggtca caggggttga gtttttccga gttggtcgaa
     2221 gggtgatgtc agtgcttggg accttggtgg ttgtacttgt gtattggctg atgtcattga
     2281 ggttatgcaa gtggggcacc agctcgagta agataccggt acctccggcc agaccggccg
     2341 ctttatacag ctcatccata cccagggtaa taccggcagc ggtcacaaac tccagcagca
     2401 ccatgtggtc gcgcttctcg ttcggatctt tgctcagttt gctttggtaa gacagataat
     2461 ggttatccgg caacaggacc ggaccgtcac caatcggggt gttttgctgg taatgatccg
     2521 ccagttgaac accaccgtcc tcaatgttgt gacggatttt gaaattcgct ttgatgccat
     2581 ttttctgttt atctgcggtg atgtaaacgt tgtgggaatt gtaattgtac tccagcttgt
     2641 gacccaggat attgccatcc tctttgaaat caataccctt cagctcaatg cgattcacca
     2701 gggtgtcgcc ttcgaatttg acttcggcac gcgtcttgta gttaccgtcg tctttgaaaa
     2761 agatcgtacg ctcctgcaca tagccttctg gcatcgcgct cttaaagaaa tcgtgttgtt
     2821 tcatgtggtc cggataacgc gcgaaacact gaacgccata acccaaggtc gtgaccaggg
     2881 taggccacgg gaccggcagc ttacccgtcg tgcaaatcag cttcagggtc agcttgccgt
     2941 aagttgcgtc accttcaccc tcaccgctca cgctaaactt atgaccgtta acgtcgccgt
     3001 ctaattcaac taagatcggt actacgcctg taaaaagttc ttcgcccttg ctaaccaggc
     3061 cagaaaggcc caggtcctcc tcgctgatca gcttctgctc gaattcgtca ccagtggaac
     3121 ctggaaccca gagcagcagt acccatagca ggagtgtgtc tgtctccatg gtggatatcc
     3181 ccaagccggt accgaattct agtataactt cgtataggat actttatacg aagttatcat
     3241 tgggattctt cctattttga tccaagcatc accatcgacc ctctagtcca gatctcacca
     3301 tcgacccata acttcgtata gcatacatta tacgaagtta tgtccctcga agaggttcga
     3361 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3421 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3481 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3541 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3601 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3661 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3721 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3781 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3841 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3901 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3961 cgcctccccg catcgatacc gtcgacctcg acccgggcgg ccgcttcgag cagacatgat
     4021 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4081 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4141 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     4201 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     4261 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4321 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgcccgg gctttgcccg
     4381 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     4441 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4501 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4561 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5461 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5521 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5581 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5641 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5701 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5761 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5821 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5881 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5941 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6001 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6061 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6121 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6181 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6241 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6301 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6361 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     6421 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     6481 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     6541 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6601 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6661 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6721 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6781 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6841 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6901 gggcctcttc gctattacgc ca
//
