LOCUS       Exported                6950 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  This AAV plasmid in the presence of Cre recombinase expresses an 
            extracellular dL5 FAP fused to the neuroligin-1 cytoplasmic domain 
            for targeting to the synapse with a cell fill dTomato..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.hSyn.FLEX.FAPdL5-POST-T2A-dTomato.FLEX.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  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 6950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6950
                     /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(2342..3061)
                     /codon_start=1
                     /product="fluorogen-activating protein composed of a dimer 
                     of the L5-MG single-chain antibody fragment (Szent-Gyorgyi 
                     et al., 2008)"
                     /label=FAP-alpha-2
                     /note="binds malachite green derivatives to produce red 
                     fluorescence"
                     /translation="QAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGHYANWFQQKPGQA
                     PRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSLSDVDGYLFGGGT
                     QLTVLSGGGGSGGGGSGGGGSGGGGSQAVVTQEPSVTVSPGGTVILTCGSGTGAVTSGH
                     YANWFQQKPGQAPRALIFDTDKKYSWTPGRFSGSLLGAKAALTISDAQPEDEAEYYCSL
                     SDVDGYLFGGGTQLTVLS"
     CDS             complement(3077..3106)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     sig_peptide     complement(3113..3175)
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     protein_bind    3210..3243
                     /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    3314..3347
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3389..3977
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3442..3462)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3860..3871)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3980..3996)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3981..3997)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4026..4147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4037..4056)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4091..4110
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4309..4438
                     /label=AAV2 ITR
     primer_bind     complement(4504..4521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4675..5263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4755..4774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5434..6294)
                     /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     6057..6076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6295..6399)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6425..6880)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6562..6583)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6774..6793
                     /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 cggagaggac ggtcagctgg gtgcctcctc cgaacagata accgtcaacg tcggagagcg
     2401 aacagtaata ctcagcctca tcttcaggct gcgcatccga gatggtcagg gcagccttgg
     2461 ccccaaggag ggagcctgag aatcggccag gggtccagga atacttcttg tcggtgtcaa
     2521 atataagtgc cctgggggct tggccaggct tctgctggaa ccagttggca taatgaccac
     2581 tggtgacagc tccagtgccg gagccacaag tgagaatgac tgtccctcct ggggacacag
     2641 tcactgacgg ctcctgagtc accacagcct gagaaccgcc gcctccggaa ccaccaccgc
     2701 cgctgccacc gccaccagag ccgccaccac cggacagaac cgtcagttgt gttccaccgc
     2761 caaacaagta gccatccacg tcgctaagag aacagtaata ctcggcttcg tcctcaggct
     2821 gagcatctga tattgtcaaa gctgcctttg ctcccagcaa agatccagaa aatctacctg
     2881 gggtccaaga atacttctta tcagtatcaa aaatcaaagc tcttggagct tgtcctggtt
     2941 tttgttgaaa ccaatttgca taatgaccag atgtgacggc acccgtacca cttccacaag
     3001 taagtataac agtaccacct gggctaacgg taacactagg ttcttgggta acgacggcct
     3061 gcaggccaga aaggcccagg tcctcctcgc tgatcagctt ctgctcgaat tcgtcaccag
     3121 tggaacctgg aacccagagc agcagtaccc atagcaggag tgtgtctgtc tccatggtgg
     3181 atatccccaa gccggtaccg aattctagta taacttcgta taggatactt tatacgaagt
     3241 tatcattggg attcttccta ttttgatcca agcatcacca tcgaccctct agtccagatc
     3301 tcaccatcga cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag
     3361 gttcgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     3421 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3481 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3541 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3601 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3661 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3721 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3781 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3841 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     3901 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     3961 ttgggccgcc tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga
     4021 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4081 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4141 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4201 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     4261 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     4321 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4381 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4441 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5101 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5341 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5401 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5461 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5521 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5581 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5641 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5701 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5761 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5821 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5881 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5941 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6001 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6061 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6121 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6181 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6241 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6301 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6361 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6421 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6481 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6541 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6601 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6661 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6721 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6781 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6841 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     6901 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
//
