LOCUS       sequence-322444-        5968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the full length TurboID on the surface of astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5968)
  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 5968)
  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: pZac2.1-GfaABC1D-TurboID(full)-HA-GPI
FEATURES             Location/Qualifiers
     source          1..5968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1451..1460
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1538..2494
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             2498..2524
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2582..2591
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2993..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3004..3023)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3058..3077
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3276..3405
                     /label=AAV2 ITR
     primer_bind     complement(3424..3441)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3424..3440)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3433..3455)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3582..4037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3669..3688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3879..3900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4063..4167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4168..4236,4237..5028)
                     /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"
     rep_origin      5199..5787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5688..5707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     1441 tcgagaattc gccaccatgg tagagatgct gccaactgtc gctgtgctgg tcttggcagt
     1501 gtccgtggtt gccaaggata acaccacgcg tttaaacaaa gacaatactg tgcctctgaa
     1561 gctgatcgct ctcctggcta atggcgagtt ccatagtggc gaacagctgg gagaaaccct
     1621 gggcatgtcc agggccgcta tcaacaagca cattcagact ctgcgcgact ggggcgtgga
     1681 cgtgttcacc gtgcccggaa agggctactc tctgcccgag cctatcccgc tgctgaacgc
     1741 taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg tggtcgactc
     1801 caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg cttgcattgc
     1861 agaatatcag caggcaggga gaggaagcag agggaggaaa tggttctctc cttttggagc
     1921 taacctgtac ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa tcggcctggg
     1981 cccggtcatc ggaattgtca tggcagaagc gctgcgaaag ctgggagcag acaaggtgcg
     2041 agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca tcctggtgga
     2101 gctggccgga ataacaggcg atgctgcaca gatcgtcatt ggcgccggga ttaacgtggc
     2161 tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc aggaagcagg
     2221 gattaacctg gacaggaata ctctggccgc tacgctgatc cgagagctgc gggcagccct
     2281 ggaactgttc gagcaggaag gcctggctcc atatctgcca cggtgggaga agctggataa
     2341 cttcatcaat agacccgtga agctgatcat tggggacaaa gagattttcg ggattagccg
     2401 ggggattgat aaacagggag ccctgctgct ggaacaggac ggagttatca aaccctggat
     2461 gggcggagaa atcagtctgc ggtctgccga aaagctttac ccttacgatg taccggatta
     2521 cgcaactagt ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcggctag
     2581 cgccaccatg gcccagctca agaagaagct gcaagccctg aagaagaaga acgcccagct
     2641 caagtggaag ctccaggccc tgaagaagaa gctggcccag aaggctagcg gtggaggcgg
     2701 ttcaggcgga ggtggctctg gcggtggcgg atcgtctagc ggtggaggcg gttcaggcgg
     2761 aggtggctct ggcggtggcg gatcgtctag cggtggaggc ggttcaggcg gaggtggctc
     2821 tggcggtggc ggatcgtcta gctaccaact cgagaatggc ggcataagcc tgctggttca
     2881 gaacacatcc tggatgctgc tgctgctgct ttccctctcc ctcctccaag ccctggactt
     2941 catttctctg tagtctagag tcgacccggg cggccgcttc gagcagacat gataagatac
     3001 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3061 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3121 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     3181 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     3241 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     3301 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3361 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta attaacctaa
     3421 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3481 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3541 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     3601 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3661 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3721 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3781 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3841 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3901 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3961 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4021 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4141 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4201 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4261 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4321 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4381 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4441 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4501 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4561 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4621 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4681 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4741 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4801 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4861 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4921 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4981 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5041 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5881 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5941 agcgagtcag tgagcgagga agcggaag
//
