LOCUS       sequence-327742-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses c-terminal half of TurboID on surface of astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5184)
  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 5184)
  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(C )-HA-GPI
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    115..136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(151..156,157..174,175..181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    189..205
                     /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     194..216
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     213..229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     213..229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     intron          1225..1357
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1340..1359
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1402..1420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1437..1446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1722..1748
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1806..1815
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2217..2338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2228..2247)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2282..2301
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2500..2629
                     /label=AAV2 ITR
     primer_bind     complement(2648..2665)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2648..2664)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2657..2679)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2806..3261
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2893..2912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3103..3124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3287..3391
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3392..3460,3461..4252)
                     /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      4423..5011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4912..4931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      121 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      181 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      241 cagatttaat taaggcctta attaggctgc gcgctcgctc gctcactgag gccgcccggg
      301 caaagcccgg gcggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      361 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
      421 ctctaggaag atctaacata tcctggtgtg gagtagggga cgctgctctg acagaggctc
      481 gggggcctga gctggctctg tgagctgggg aggaggcaga cagccaggcc ttgtctgcaa
      541 gcagacctgg cagcattggg ctggccgccc cccagggcct cctcttcatg cccagtgaat
      601 gactcacctt ggcacagaca caatgttcgg ggtgggcaca gtgcctgctt cccgccgcac
      661 cccagccccc ctcaaatgcc ttccgagaag cccattgagc agggggcttg cattgcaccc
      721 cagcctgaca gcctggcatc ttgggataaa agcagcacag ccccctaggg gctgcccttg
      781 ctgtgtggcg ccaccggcgg tggagaacaa ggctctattc agcctgtgcc caggaaaggg
      841 gatcagggga tgcccaggca tggacagtgg gtggcagggg gggagaggag ggctgtctgc
      901 ttcccagaag tccaaggaca caaatgggtg aggggagagc tctccccata gctgggctgc
      961 ggcccaaccc caccccctca ggctatgcca gggggtgttg ccaggggcac ccgggcatcg
     1021 ccagtctagc ccactccttc ataaagccct cgcatcccag gagcgagcag agccagagca
     1081 ggttggagag gagacgcatc acctccgctg ctcgcaagct ttattgcggt agtttatcac
     1141 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcaga
     1201 agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac
     1261 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt
     1321 ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac
     1381 agctcttaag gctagagtac ttaatacgac tcactatagg ctagcctcga gaattcgcca
     1441 ccatggtaga gatgctgcca actgtcgctg tgctggtctt ggcagtgtcc gtggttgcca
     1501 aggataacac cacgcgttta aacggcctgg ctccatatct gccacggtgg gagaagctgg
     1561 ataacttcat caatagaccc gtgaagctga tcattgggga caaagagatt ttcgggatta
     1621 gccgggggat tgataaacag ggagccctgc tgctggaaca ggacggagtt atcaaaccct
     1681 ggatgggcgg agaaatcagt ctgcggtctg ccgaaaagct ttacccttac gatgtaccgg
     1741 attacgcaac tagtggtgga ggcggttcag gcggaggtgg ctctggcggt ggcggatcgg
     1801 ctagcgccac catggcccag ctcaagaaga agctgcaagc cctgaagaag aagaacgccc
     1861 agctcaagtg gaagctccag gccctgaaga agaagctggc ccagaaggct agcggtggag
     1921 gcggttcagg cggaggtggc tctggcggtg gcggatcgtc tagcggtgga ggcggttcag
     1981 gcggaggtgg ctctggcggt ggcggatcgt ctagcggtgg aggcggttca ggcggaggtg
     2041 gctctggcgg tggcggatcg tctagctacc aactcgagaa tggcggcata agcctgctgg
     2101 ttcagaacac atcctggatg ctgctgctgc tgctttccct ctccctcctc caagccctgg
     2161 acttcatttc tctgtaatct agagtcgacc cgggcggccg cttcgagcag acatgataag
     2221 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2281 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     2341 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     2401 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     2461 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     2521 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2581 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac
     2641 ctaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     2701 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     2761 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     2821 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2881 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2941 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3001 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3061 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3121 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3181 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3241 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3301 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3361 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3421 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3481 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3541 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3601 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3661 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3721 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3781 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3841 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3901 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3961 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4021 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4081 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4141 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4201 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4261 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4321 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4381 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4441 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4501 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4561 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4621 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4681 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4741 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4801 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4861 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4921 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4981 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5041 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5101 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5161 gcgcagcgag tcagtgagcg agga
//
