LOCUS       sequence-348789-        8359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Glt-1 fused with BioID2 at the C-terminus in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8359)
  AUTHORS   Soto JS, Jami-Alahmadi Y, Chacon J, Moye SL, Diaz-Castro B, 
            Wohlschlegel JA, Khakh BS
  TITLE     Astrocyte-neuron subproteomes and obsessive-compulsive disorder 
            mechanisms.
  JOURNAL   Nature. 2023 Apr;616(7958):764-773. doi: 10.1038/s41586-023-05927-7.
            Epub 2023 Apr 12.
  PUBMED    37046092
REFERENCE   2  (bases 1 to 8359)
  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/s41586-023-05927-7"; journalName: "Nature"; date: 
            "2023-04"; volume: "616"; issue: "7958"; pages: "764-773"
COMMENT     Sequence Label: pZac2.1 gfaABC1D-Glt1-BioID2-BioID2-HA
FEATURES             Location/Qualifiers
     source          1..8359
                     /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      1447..1456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3190..3888
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="MFKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKW
                     LSQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKV
                     SGVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISE
                     NLKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKE
                     ILSGEFSLRRS"
     CDS             3973..4671
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="MFKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKW
                     LSQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKV
                     SGVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISE
                     NLKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKE
                     ILSGEFSLRRS"
     CDS             4681..4707
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4732..5320
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4785..4805)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5203..5214)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5384..5505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5395..5414)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5449..5468
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5667..5796
                     /label=AAV2 ITR
     primer_bind     complement(5815..5832)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5815..5831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5824..5846)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5973..6428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6060..6079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6270..6291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6454..6558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6559..6627,6628..7419)
                     /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      7590..8178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8079..8098
                     /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 tcgaacgcca ccatggcatc aacagagggt gccaacaata tgcccaagca ggtagaagtg
     1501 cgcatgcatg acagccacct cagctccgat gagccaaagc accgaaacct gggcatgcgc
     1561 atgtgcgaca agctggggaa aaatctcctg ctctcactga ctgtgtttgg tgtcatcctg
     1621 ggagcagtgt gtggcgggct gcttcgcttg gcatcgccca tccaccctga tgtggtcatg
     1681 ttgatagcct tcccggggga catactcatg aggatgctga agatgctcat cctccctctt
     1741 atcatctcca gtttaatcac agggttgtca ggcctggatg ctaaagccag cggccgccta
     1801 ggcacgagag ctatggtgta ttacatgtcc acgaccatca ttgccgccgt gctgggggtc
     1861 atcctggtgt tggccatcca cccaggcaat cccaaactca agaagcagct agggcccggg
     1921 aagaagaacg acgaggtgtc tagcctggat gccttcctgg atctcattag aaatctcttc
     1981 ccggagaacc tggtgcaagc ctgtttccag cagattcaga cagtgacaaa gaaagttctg
     2041 gtggcacctc catctgagga ggccaatacc accaaggcgg tcatctccat gttgaatgaa
     2101 accatgaacg aggcccctga agaaactaag atcgttatca agaagggcct ggagttcaag
     2161 gacgggatga acgtcttagg tctgatcgga ttctttattg ctttcggcat tgccatgggg
     2221 aagatgggtg aacaggccaa gctgatggtg gagttcttca acattctgaa tgagatcgtg
     2281 atgaagttag tgatcatgat catgtggtac tcccctctgg gtatcgcctg cttgatttgt
     2341 gggaagatca tcgccatcaa ggacttagaa gtggttgcta ggcagctggg gatgtacatg
     2401 atcaccgtga tcgtgggcct catcattcac gggggcatct ttctcccctt gatttacttt
     2461 gtagtgacca gaaaaaatcc attctccttt tttgctggca tattccaagc ctggatcact
     2521 gctctgggaa ctgcttccag tgctggaact ttgcctgtta ccttccgttg cttggaagat
     2581 aatctaggga ttgacaagcg tgtgaccaga ttcgtcctcc cagtcggagc aaccattcac
     2641 atggatggca cagcccttta cgaggctgtg gcagccatct tcatagccca aatgaatggg
     2701 gtcatcttgg atggaggtca gattgtgact gtaagcctta cagccaccct ggcgagcatt
     2761 ggtgcagcca gtattcccag cgccgggctg gtcaccatgc tcctcattct cacagctgtg
     2821 ggcctgccaa cggaggatat cagtctgctg gtggcggtgg actggctgct ggatagaatg
     2881 agaacttcag tcaatgtggt gggcgattct tttggggctg ggattgtcta tcacctttcc
     2941 aagtctgagc tggacaccat tgactcccaa caccgaatgc aggaagacat cgagatgacc
     3001 aagacgcagt ccatttacga cgacaagaac cacagggaaa gcaactctaa tcagtgtgtc
     3061 tatgccgcac acaactctgt cgtaatagat gagtgcaagg taactctggc ggccaatgga
     3121 aagtcagctg actgcagtgt tgaggaagaa ccttggaaac gtgaaaaact cgagctcgga
     3181 tccaccacca tgttcaaaaa tcttatttgg ctgaaggaag ttgatagcac ccaggagcgg
     3241 cttaaggaat ggaatgtctc ctatggcact gcactcgtcg ccgatagaca gaccaaaggt
     3301 agaggcggtc tcggacggaa gtggttgagt caggagggcg gtctttactt ctctttcttg
     3361 ttgaatccaa aagagtttga aaatttgttg caacttccac tggtcttggg actctccgtg
     3421 agcgaggcac tcgaagagat tacagagata ccatttagtc tgaagtggcc caacgacgtt
     3481 tattttcagg agaaaaaggt ttcaggggtc ctttgtgagc tcagtaagga caagctgata
     3541 gtgggaatcg gtataaacgt gaatcaaaga gagatccctg aagagatcaa ggatcgggcc
     3601 acaacacttt atgaaatcac tggaaaggat tgggacagaa aggaggtctt gctgaaggtt
     3661 ctcaagcgaa tttccgaaaa tctcaagaaa tttaaggaaa aatcctttaa agagttcaaa
     3721 ggaaaaattg agtcaaagat gttgtacctt ggagaggagg ttaaactctt gggagaaggc
     3781 aagattaccg ggaaacttgt gggcctgagt gaaaagggcg gtgctttgat acttacagag
     3841 gaaggcataa aggagatact gtccggcgaa tttagcttgc gacggtcaag cttaggtgga
     3901 ggaggttctg gaggcggtgg aagtggtggc ggaggtagcg gtggaggagg ttctatggtg
     3961 tctaaggaat tcatgttcaa aaatcttatt tggctgaagg aagttgatag cacccaggag
     4021 cggcttaagg aatggaatgt ctcctatggc actgcactcg tcgccgatag acagaccaaa
     4081 ggtagaggcg gtctcggacg gaagtggttg agtcaggagg gcggtcttta cttctctttc
     4141 ttgttgaatc caaaagagtt tgaaaatttg ttgcaacttc cactggtctt gggactctcc
     4201 gtgagcgagg cactcgaaga gattacagag ataccattta gtctgaagtg gcccaacgac
     4261 gtttattttc aggagaaaaa ggtttcaggg gtcctttgtg agctcagtaa ggacaagctg
     4321 atagtgggaa tcggtataaa cgtgaatcaa agagagatcc ctgaagagat caaggatcgg
     4381 gccacaacac tttatgaaat cactggaaag gattgggaca gaaaggaggt cttgctgaag
     4441 gttctcaagc gaatttccga aaatctcaag aaatttaagg aaaaatcctt taaagagttc
     4501 aaaggaaaaa ttgagtcaaa gatgttgtac cttggagagg aggttaaact cttgggagaa
     4561 ggcaagatta ccgggaaact tgtgggcctg agtgaaaagg gcggtgcttt gatacttaca
     4621 gaggaaggca taaaggagat actgtccggc gaatttagct tgcgacggtc agctagcgct
     4681 tacccttacg atgtaccgga ttacgcatag ggatccaatt cgcttatcga taatcaacct
     4741 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     4801 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     4861 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     4921 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     4981 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5041 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5101 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     5161 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5221 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5281 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg gaattctgca
     5341 gatatctaga gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     5401 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     5461 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5521 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     5581 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     5641 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     5701 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     5761 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     5821 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5881 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5941 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     6001 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     6061 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6121 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6181 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6241 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6301 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     6361 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     6421 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6481 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6541 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6601 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6661 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6721 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6781 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6841 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6901 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6961 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7021 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7081 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7141 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7201 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7261 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7321 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7381 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7441 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7501 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7561 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7621 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7681 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7741 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7801 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7861 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7921 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7981 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8041 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8101 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8161 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8221 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8281 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8341 gtgagcgagg aagcggaag
//
