LOCUS       sequence-327746-        5574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses n-terminal half of TurboID on surface of neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  AUTHORS   Takano T, Wallace JT, Baldwin KT, Purkey AM, Uezu A, Courtland JL, 
            Soderblom EJ, Shimogori T, Maness PF, Eroglu C, Soderling SH
  TITLE     Chemico-genetic discovery of astrocytic control of inhibition in 
            vivo.
  JOURNAL   Nature. 2020 Dec;588(7837):296-302. doi: 10.1038/s41586-020-2926-0. 
            Epub 2020 Nov 11.
  PUBMED    33177716
REFERENCE   2  (bases 1 to 5574)
  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/s41586-020-2926-0"; journalName: "Nature"; date: "2020-12";
            volume: "588"; issue: "7837"; pages: "296-302"
COMMENT     Sequence Label: AAV-hSynI-V5-TurboID(N)-GPI
FEATURES             Location/Qualifiers
     source          1..5574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(264..368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     436..454
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(492..514)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     614..633
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(650..1105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(787..808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     999..1018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1180..1320
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1191..1320
                     /label=AAV2 ITR
     promoter        1357..1804
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1833..1842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1920..1961
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      2787..2796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2850..2861
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     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"
     polyA_signal    3794..4018
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4027..4156
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4218..4806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4298..4317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4977..5574,1..194,195..263))
                     /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"
ORIGIN
        1 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
       61 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
      121 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
      181 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
      241 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
      301 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
      361 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
      421 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
      481 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
      541 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
      601 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac
      661 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      721 tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt
      781 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
      841 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt
      901 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
      961 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     1021 ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     1081 cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac
     1141 cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc
     1201 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     1261 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     1321 gcggccgtgt ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag
     1381 gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc
     1441 aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg
     1501 gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg
     1561 cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc
     1621 gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc
     1681 gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg
     1741 cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc
     1801 gcagtcgaga aaccggctag agtcgaaagc ttgccaccat ggtagagatg ctgccaactg
     1861 tcgctgtgct ggtcttggca gtgtccgtgg ttgccaagga taacaccacg cgtttaaacg
     1921 gcaagcccat ccccaacccc ctgctgggcc tggacagcac cgctagcaaa gacaatactg
     1981 tgcctctgaa gctgatcgct ctcctggcta atggcgagtt ccatagtggc gaacagctgg
     2041 gagaaaccct gggcatgtcc agggccgcta tcaacaagca cattcagact ctgcgcgact
     2101 ggggcgtgga cgtgttcacc gtgcccggaa agggctactc tctgcccgag cctatcccgc
     2161 tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg
     2221 tggtcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg
     2281 cttgcattgc agaatatcag caggcaggga gaggaagcag agggaggaaa tggttctctc
     2341 cttttggagc taacctgtac ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa
     2401 tcggcctggg cccggtcatc ggaattgtca tggcagaagc gctgcgaaag ctgggagcag
     2461 acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca
     2521 tcctggtgga gctggccgga ataacaggcg atgctgcaca gatcgtcatt ggcgccggga
     2581 ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc
     2641 aggaagcagg gattaacctg gacaggaata ctctggccgc tacgctgatc cgagagctgc
     2701 gggcagccct ggaactgttc gagcaggaaa ctagtggtgg aggcggttca ggcggaggtg
     2761 gctctggcgg tggcggatcg gctagcgcca ccatggccca gctcgaaaag gagctgcaag
     2821 ccctggagaa ggagaacgcc cagctcgaat gggagctcca ggccctggag aaggagctgg
     2881 cccagaaggc tagcggtgga ggcggttcag gcggaggtgg ctctggcggt ggcggatcgt
     2941 ctagcggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatcg tctagcggtg
     3001 gaggcggttc aggcggaggt ggctctggcg gtggcggatc gtctagctac caactcgaga
     3061 atggcggcat aagcctgctg gttcagaaca catcctggat gctgctgctg ctgctttccc
     3121 tctccctcct ccaagccctg gacttcattt ctctgtaagg atccaattcg 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 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3661 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3721 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgct
     3781 cgagagatct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3841 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3901 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3961 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatgggc
     4021 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4081 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
     4141 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4201 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4261 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4321 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4381 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4441 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4501 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4561 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4621 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4681 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4741 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4801 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4861 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4921 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4981 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5041 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5101 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5161 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5221 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5281 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5341 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5401 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5461 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5521 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagt
//
