LOCUS       sequence-330209-        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    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   153..282
                     /label=AAV2 ITR
     intron          1133..1265
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1248..1267
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1310..1328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1345..1354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1432..2388
                     /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             2392..2418
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2476..2485
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2887..3008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2898..2917)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2952..2971
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3170..3299
                     /label=AAV2 ITR
     primer_bind     complement(3318..3335)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3318..3334)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3327..3349)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3476..3931
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3563..3582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3773..3794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3957..4061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4062..4130,4131..4922)
                     /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      5093..5681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5582..5601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaga tttaattaag gccttaatta ggctgcgcgc tcgctcgctc actgaggccg
      181 cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag
      241 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
      301 cgccatgcta cttatctacg tagccatgct ctaggaagat ctaacatatc ctggtgtgga
      361 gtaggggacg ctgctctgac agaggctcgg gggcctgagc tggctctgtg agctggggag
      421 gaggcagaca gccaggcctt gtctgcaagc agacctggca gcattgggct ggccgccccc
      481 cagggcctcc tcttcatgcc cagtgaatga ctcaccttgg cacagacaca atgttcgggg
      541 tgggcacagt gcctgcttcc cgccgcaccc cagcccccct caaatgcctt ccgagaagcc
      601 cattgagcag ggggcttgca ttgcacccca gcctgacagc ctggcatctt gggataaaag
      661 cagcacagcc ccctaggggc tgcccttgct gtgtggcgcc accggcggtg gagaacaagg
      721 ctctattcag cctgtgccca ggaaagggga tcaggggatg cccaggcatg gacagtgggt
      781 ggcagggggg gagaggaggg ctgtctgctt cccagaagtc caaggacaca aatgggtgag
      841 gggagagctc tccccatagc tgggctgcgg cccaacccca ccccctcagg ctatgccagg
      901 gggtgttgcc aggggcaccc gggcatcgcc agtctagccc actccttcat aaagccctcg
      961 catcccagga gcgagcagag ccagagcagg ttggagagga gacgcatcac ctccgctgct
     1021 cgcaagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca gtgcttctga
     1081 cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat
     1141 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     1201 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     1261 cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc
     1321 actataggct agcctcgaga attcgccacc atggtagaga tgctgccaac tgtcgctgtg
     1381 ctggtcttgg cagtgtccgt ggttgccaag gataacacca cgcgtttaaa caaagacaat
     1441 actgtgcctc tgaagctgat cgctctcctg gctaatggcg agttccatag tggcgaacag
     1501 ctgggagaaa ccctgggcat gtccagggcc gctatcaaca agcacattca gactctgcgc
     1561 gactggggcg tggacgtgtt caccgtgccc ggaaagggct actctctgcc cgagcctatc
     1621 ccgctgctga acgctaaaca gattctggga cagctggacg gcgggagcgt ggcagtcctg
     1681 cctgtggtcg actccaccaa tcagtacctg ctggatcgaa tcggcgagct gaagagtggg
     1741 gatgcttgca ttgcagaata tcagcaggca gggagaggaa gcagagggag gaaatggttc
     1801 tctccttttg gagctaacct gtacctgagt atgttttggc gcctgaagcg gggaccagca
     1861 gcaatcggcc tgggcccggt catcggaatt gtcatggcag aagcgctgcg aaagctggga
     1921 gcagacaagg tgcgagtcaa atggcccaat gacctgtatc tgcaggatag aaagctggca
     1981 ggcatcctgg tggagctggc cggaataaca ggcgatgctg cacagatcgt cattggcgcc
     2041 gggattaacg tggctatgag gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca
     2101 ctgcaggaag cagggattaa cctggacagg aatactctgg ccgctacgct gatccgagag
     2161 ctgcgggcag ccctggaact gttcgagcag gaaggcctgg ctccatatct gccacggtgg
     2221 gagaagctgg ataacttcat caatagaccc gtgaagctga tcattgggga caaagagatt
     2281 ttcgggatta gccgggggat tgataaacag ggagccctgc tgctggaaca ggacggagtt
     2341 atcaaaccct ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagct ttacccttac
     2401 gatgtaccgg attacgcaac tagtggtgga ggcggttcag gcggaggtgg ctctggcggt
     2461 ggcggatcgg ctagcgccac catggcccag ctcaagaaga agctgcaagc cctgaagaag
     2521 aagaacgccc agctcaagtg gaagctccag gccctgaaga agaagctggc ccagaaggct
     2581 agcggtggag gcggttcagg cggaggtggc tctggcggtg gcggatcgtc tagcggtgga
     2641 ggcggttcag gcggaggtgg ctctggcggt ggcggatcgt ctagcggtgg aggcggttca
     2701 ggcggaggtg gctctggcgg tggcggatcg tctagctacc aactcgagaa tggcggcata
     2761 agcctgctgg ttcagaacac atcctggatg ctgctgctgc tgctttccct ctccctcctc
     2821 caagccctgg acttcatttc tctgtaatct agagtcgacc cgggcggccg cttcgagcag
     2881 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2941 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3001 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3061 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     3121 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3181 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3241 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3301 cttaattaac ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3361 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3421 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     3481 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3541 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3601 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3661 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3721 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3781 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3841 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3901 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     3961 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4021 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4081 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4141 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4201 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4261 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4321 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4381 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4441 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4501 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4561 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4621 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4681 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4741 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4801 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4861 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4921 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4981 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5041 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5101 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5161 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5221 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5281 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5341 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5401 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5461 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5521 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5581 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5641 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5701 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5761 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5821 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5881 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5941 ggaaagcggg cagtgagcgc aacgcaat
//
