LOCUS       Exported                6806 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  This AAV plasmid constitutively 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.FAPdL5-POST-T2A-dTomato.WPRE.bGH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  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 6806)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6806
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    175..223
                     /note="synthetic polyadenylation signal"
     misc_feature    237..328
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     277..296
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        364..811
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     sig_peptide     857..919
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     CDS             926..955
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             971..1690
                     /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(1988..1999)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2396..2449
                     /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"
     misc_feature    3180..3768
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3233..3253)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3651..3662)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3801..3818)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3807..4014
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4023..4152
                     /label=AAV2 ITR
     rep_origin      4223..4678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4310..4329)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4520..4541
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4745..4764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4864..4886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4924..4942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5010..5114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5115..5975
                     /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(5333..5352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6146..6734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6635..6654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6796..119
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 cggccgcacg cgttctagaa atattaaggt acgggaggta cttggagcgg ccgcaataaa
      181 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
      241 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
      301 gcaagtgcag gtgccagaac atttctctat tctagactcg agctgcagag ggccctgcgt
      361 atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac
      421 cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc
      481 agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc
      541 gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg
      601 cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt
      661 ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg
      721 gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag
      781 cggaggagtc gtgtcgtgcc tgagagcgca gtcgaattca agctgctagc aaggatccgg
      841 cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
      901 tccaggttcc actggtgacg aattcgagca gaagctgatc agcgaggagg acctgggcct
      961 ttctggcctg caggccgtcg ttacccaaga acctagtgtt accgttagcc caggtggtac
     1021 tgttatactt acttgtggaa gtggtacggg tgccgtcaca tctggtcatt atgcaaattg
     1081 gtttcaacaa aaaccaggac aagctccaag agctttgatt tttgatactg ataagaagta
     1141 ttcttggacc ccaggtagat tttctggatc tttgctggga gcaaaggcag ctttgacaat
     1201 atcagatgct cagcctgagg acgaagccga gtattactgt tctcttagcg acgtggatgg
     1261 ctacttgttt ggcggtggaa cacaactgac ggttctgtcc ggtggtggcg gctctggtgg
     1321 cggtggcagc ggcggtggtg gttccggagg cggcggttct caggctgtgg tgactcagga
     1381 gccgtcagtg actgtgtccc caggagggac agtcattctc acttgtggct ccggcactgg
     1441 agctgtcacc agtggtcatt atgccaactg gttccagcag aagcctggcc aagcccccag
     1501 ggcacttata tttgacaccg acaagaagta ttcctggacc cctggccgat tctcaggctc
     1561 cctccttggg gccaaggctg ccctgaccat ctcggatgcg cagcctgaag atgaggctga
     1621 gtattactgt tcgctctccg acgttgacgg ttatctgttc ggaggaggca cccagctgac
     1681 cgtcctctcc gcggccggtc tggccggagg taccggtatc ttactcgagc tggtgcccca
     1741 cttgcataac ctcaatgaca tcagccaata cacaagtaca accaccaagg tcccaagcac
     1801 tgacatcacc cttcgaccaa ctcggaaaaa ctcaacccct gtgaccagcg catttccaac
     1861 cgctaagcaa gacgacccca aacaacagcc tagtccattt agcgtagacc aaagggacta
     1921 cagcacagag ctgtccgtga ccatagctgt aggagcttcc ctgcttttcc tgaacattct
     1981 ggctttcgcg gccctctatt acaagaagga caaaaggcgc cacgacgtac ataggagatg
     2041 tagtccgcaa cgaactacaa ccaatgatct tacgcacgca cctgaggagg agatcatgag
     2101 tcttcagatg aaacacacag acctggacca cgagtgtgaa tccatccatc cccatgaggt
     2161 ggtgctgagg acagcatgcc ctccggacta taccctggca atgcgccgaa gcccagacga
     2221 cattcccctg atgacaccca ataccataac catgattccc aacactattc caggaatcca
     2281 gcctctccac actttcaaca cattcactgg tggacagaat aatactctcc ctcatccgca
     2341 tccacacccc cactcccaca gcacaacacg ggtgggtggc agcggtggca gatctgaggg
     2401 tcggggctct ctgctcacat gtggcgacgt cgaggagaat cccggaccgg ccccggggtc
     2461 gacaatggtg tctaagggtg aggaagtaat taaggagttt atgagattca aagttagaat
     2521 ggagggtagc atgaacgggc acgagtttga aattgaaggt gagggtgaag gaagaccata
     2581 tgagggcaca caaaccgcca agctcaaggt taccaaaggt ggcccccttc ctttcgcctg
     2641 ggacatcctc agccctcaat tcatgtacgg tagcaaggct tacgtcaaac accctgctga
     2701 tatacctgat tacaaaaaac tgtcatttcc tgaaggattt aagtgggaac gagtaatgaa
     2761 ttttgaggat ggtgggctgg tcactgtcac gcaagactcc tctctgcagg acggcaccct
     2821 gatttacaag gtgaagatgc gcggaacaaa ttttcccccc gatggccccg tgatgcagaa
     2881 gaagaccatg gggtgggagg cctctacaga acgactgtac ccccgcgatg gcgttcttaa
     2941 aggcgaaatc catcaggcgc ttaaactcaa agatggcggg cactacctgg ttgagttcaa
     3001 aactatatat atggccaaaa agccggttca gctgcctggc tactactacg tggacaccaa
     3061 gttggacatc acgagtcaca acgaggacta cacaatcgtc gagcagtacg aacgctcaga
     3121 aggtcggcat cacctgttct tgtatggcat ggatgagctg tataagtaag cttatcgata
     3181 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3241 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3301 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3361 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3421 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3481 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3541 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3601 cctgtgttgc cacctggatt ctgcgcggga cgcccttctg ctacgtccct tcggccctca
     3661 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3721 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3781 gactagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3841 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3901 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3961 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggagagatc
     4021 tgaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4081 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4141 gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4201 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4261 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4321 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4381 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4441 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4501 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4561 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4621 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     4681 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     4741 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4801 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4861 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4921 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4981 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5041 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5101 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5161 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5221 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5281 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5341 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5401 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5461 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5521 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5581 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5641 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5701 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5761 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5821 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5881 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5941 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6001 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6781 cttttgctca catgtcctgc aggcag
//
