LOCUS       sequence-348844-        8769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SAPAP3 protein fused with BioID2 at the N-terminus in 
            astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8769)
  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 8769)
  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-HA-BioID2-SAPAP3
FEATURES             Location/Qualifiers
     source          1..8769
                     /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"
     CDS             1438..1464
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1474..2172
                     /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             2851..2880
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2854..2883
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2857..2886
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    5142..5730
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5195..5215)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5613..5624)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5794..5915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5805..5824)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5859..5878
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6077..6206
                     /label=AAV2 ITR
     primer_bind     complement(6225..6242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6225..6241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6234..6256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6383..6838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6470..6489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6680..6701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6864..6968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6969..7037,7038..7829)
                     /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      8000..8588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8489..8508
                     /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 taggatgtac
     1441 ccttacgatg taccggatta cgcagctagc accatgttca aaaatcttat ttggctgaag
     1501 gaagttgata gcacccagga gcggcttaag gaatggaatg tctcctatgg cactgcactc
     1561 gtcgccgata gacagaccaa aggtagaggc ggtctcggac ggaagtggtt gagtcaggag
     1621 ggcggtcttt acttctcttt cttgttgaat ccaaaagagt ttgaaaattt gttgcaactt
     1681 ccactggtct tgggactctc cgtgagcgag gcactcgaag agattacaga gataccattt
     1741 agtctgaagt ggcccaacga cgtttatttt caggagaaaa aggtttcagg ggtcctttgt
     1801 gagctcagta aggacaagct gatagtggga atcggtataa acgtgaatca aagagagatc
     1861 cctgaagaga tcaaggatcg ggccacaaca ctttatgaaa tcactggaaa ggattgggac
     1921 agaaaggagg tcttgctgaa ggttctcaag cgaatttccg aaaatctcaa gaaatttaag
     1981 gaaaaatcct ttaaagagtt caaaggaaaa attgagtcaa agatgttgta ccttggagag
     2041 gaggttaaac tcttgggaga aggcaagatt accgggaaac ttgtgggcct gagtgaaaag
     2101 ggcggtgctt tgatacttac agaggaaggc ataaaggaga tactgtccgg cgaatttagc
     2161 ttgcgacggt cacctcgagc tcggatcatg aggggttacc atggcgaccg aggcagccat
     2221 ccccgcccag cccgctttgc tgaccaacag catatggacg tgggcccagc tgccagggcc
     2281 ccgtacctgc tgggctccag ggaggccttc tccaccgagc cccgcttctg tgctccgaga
     2341 gctggcctgg gacacctttc tccagaagga cccctgagcc tgagtgaggg tccatcatca
     2401 gtaggccctg agggaggccc agggggggtg ggggctgggg gaggcagcag taccttcccc
     2461 aggatgtacc ccggccaggg ccccttcgac acctgtgaag actgtgtggg ccacccacag
     2521 ggcaagggtg ccacccgact gcctcccaca ctcctggacc agtttgaaaa gcaattgcca
     2581 gtccaacaag atggcttcca cacactccca taccagcgag gtccagcagg gcctggtcct
     2641 ggacctggat ctggcgctgc ccccgaggct cgcagcgaga gtcctagccg catccggcac
     2701 ctggttcact cagtgcagaa gctctttgcc aagtcccact ctctggaggc acccgggaag
     2761 cgagactata acgggcccaa ggccgacgga agaggtagct ctgggggaga cagctactct
     2821 ggtccaggct ctggaggcac ccccacctcc caccaccacc atcaccacca ccatcaccac
     2881 caccaccagt cccggcatgg caaacggagc aagagcaagg accgcaaggg ggatgggcga
     2941 caccagacca aggccacagg ctggtggagc tctgatgaca atttggacag tgacagtggc
     3001 tttctgggtg ggagacctcc tggagagcct ggtggcccct tctgcctgga tgctccagat
     3061 gggtcctaca gggacttgag cttcaagggg cgttcgggcg ggtcggaagg ccgctgcctt
     3121 gcctgcactg gcatgtccat gtccctggat ggacagtcag tcaagcgaag tgcctggcat
     3181 accatgatgg tcagccaggg ccgggatgga tacccagggg ccgggccagg caaggggctc
     3241 ctgggtccag agaccaaggc caaagccagg acttatcact atttgcaggt gccgcaagat
     3301 gactgggggg gttaccccac gggtggcaag gatggggaga tcccctgccg caggatgcgg
     3361 agcggcagct acatcaaagc catgggggat gaggagagtg gagactcaga tggtagcccc
     3421 aagacgtctc caaaagcact tgcccgacgc tttgcctccc gccgttcctc cagcgtggac
     3481 acagccagga tcaactgctg tgtcccaccc cggatccatc cccggagttc catccctggc
     3541 tacagccgtt ccctcacaac tggacagcta agcgaagagt ttaaccagca gctggaggcc
     3601 gtgtgcgggt ccgtgtttgg ggagcttgag tcccaggccg tggacgccct ggacctgccc
     3661 ggctgtttcc gcatgcggag ccacagctac ctccgggcca tccaggccgg ctgctctcaa
     3721 gacgacgact gcctacccct ccttgctgcc cctgcctccg tctcagggag gcccggctcc
     3781 tccttcaact tcagaaaggc cccgcccccc atcccgccgg gaagccaggc cccaccccgc
     3841 atctccatca ccgcccagag cagcaccgac tctgcccacg agagcttcac cgcagcagag
     3901 ggcccggccc ggcgctgcag ctccgcggac gggctggacg gccccaccat gggcgcacgc
     3961 accctcgagt tggcaccggt gccaccccga gccagtccca agccccccac actcatcatc
     4021 aagaccatcc ctggccggga ggagctgcgg agcttggcac gacagcggaa gtggcggccg
     4081 tccattgggg tgcaggtgga gacaatctca gactcggaca ccgagaacag gagtcgaaga
     4141 gaattccact ccattggcgt tcaggtggaa gaggacaaga ggcgggcgag gttcaagcgc
     4201 tccaacagcg tgacagccgg tgtgcaggca gacctggagc tggagggcct ggccggcctg
     4261 gccactgtgg ccacagaaga caaggccctg cagttcggac gctccttcca gaggcacgcc
     4321 tctgagcccc agcccggccc tcgggccccc acctactcag tcttccgcac ggtccacaca
     4381 cagggccagt gggcctaccg cgagggctac ccactgccgt atgagccgcc ggccaccgat
     4441 gggtcgcctg gccctacccc tgtcccagcc cccggtcccg ggtccggccg ccgtgattcc
     4501 tggatggagc ggggttcacg tagcctcccc gactcaggcc gcacgtcccc ctgcccacgg
     4561 gatggcgagt ggttcatcaa gatgctgcgg gcagaggtgg agaaactgga acactggtgc
     4621 cagcagatgg agcgtgaggc ggaggactat gagctgcccg aggagatctt agagaagatt
     4681 cgcagtgccg tgggcagtac gcaacttctc ctgtctcaga aggtccagca gttcttcagg
     4741 ctgtgtcagc aaagcttgga ccccactgcg ttccctgtgc ccaccttcca ggacctggcc
     4801 ggtttctggg acctcctgca gctctccatt gaggatgtga ccctcaagtt cctggaacta
     4861 caacaactca aggccaacag ctggaaactc ctggagccga aggaggagaa gaaggtccct
     4921 ccgccgatac caaagaagcc ctcgcggggg cggggcgtgc cggtgaagga gcgctccctg
     4981 gactcggtgg accggcagcg gcaggaagcg cgcaagcggc tcctggcggc caagcgcgcc
     5041 gcctcgttcc gccacagttc ggctaccgag agcgcggaca gcatcgagat ctacatcccc
     5101 gaggcccaga ccaggctgta ggatccaatt cgcttatcga taatcaacct ctggattaca
     5161 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5221 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5281 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5341 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5401 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5461 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5521 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     5581 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5641 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5701 gtcggatctc cctttgggcc gcctccccgc atcgataccg gaattctgca gatatctaga
     5761 gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     5821 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     5881 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     5941 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat
     6001 gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     6061 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     6121 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     6181 gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta
     6241 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6301 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     6361 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6421 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6481 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6541 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6601 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6661 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6721 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6781 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     6841 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6901 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6961 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     7021 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7081 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7141 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7201 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7261 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7321 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7381 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7441 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7501 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7561 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7621 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7681 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7741 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7801 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7861 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7921 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7981 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     8041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8161 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8221 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8281 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8341 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8401 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8461 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8521 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8581 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8641 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8701 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     8761 aagcggaag
//
