LOCUS       sequence-348838-        8489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SAPAP3 protein fused with BioID2 at the N-terminus in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8489)
  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 8489)
  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 hSyn1-HA-BioID2-SAPAP3
FEATURES             Location/Qualifiers
     source          1..8489
                     /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
     promoter        465..912
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          959..1091
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1074..1093
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1136..1154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1158..1184
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1194..1892
                     /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             2571..2600
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2574..2603
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2577..2606
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    4862..5450
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4915..4935)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5333..5344)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5514..5635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5525..5544)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5579..5598
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5797..5926
                     /label=AAV2 ITR
     primer_bind     complement(5945..5962)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5945..5961)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5954..5976)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6103..6558
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6190..6209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6400..6421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6584..6688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6689..6757,6758..7549)
                     /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"
     primer_bind     complement(6907..6926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7720..8308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8209..8228
                     /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 gaagatctga gggccctgcg tatgagtgca agtgggtttt
      481 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      541 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      601 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
      661 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
      721 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
      781 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
      841 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
      901 ctgagagcgc agtcgagaga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
      961 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     1021 gagaaaactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1081 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata
     1141 cgactcacta taggatgtac ccttacgatg taccggatta cgcagctagc accatgttca
     1201 aaaatcttat ttggctgaag gaagttgata gcacccagga gcggcttaag gaatggaatg
     1261 tctcctatgg cactgcactc gtcgccgata gacagaccaa aggtagaggc ggtctcggac
     1321 ggaagtggtt gagtcaggag ggcggtcttt acttctcttt cttgttgaat ccaaaagagt
     1381 ttgaaaattt gttgcaactt ccactggtct tgggactctc cgtgagcgag gcactcgaag
     1441 agattacaga gataccattt agtctgaagt ggcccaacga cgtttatttt caggagaaaa
     1501 aggtttcagg ggtcctttgt gagctcagta aggacaagct gatagtggga atcggtataa
     1561 acgtgaatca aagagagatc cctgaagaga tcaaggatcg ggccacaaca ctttatgaaa
     1621 tcactggaaa ggattgggac agaaaggagg tcttgctgaa ggttctcaag cgaatttccg
     1681 aaaatctcaa gaaatttaag gaaaaatcct ttaaagagtt caaaggaaaa attgagtcaa
     1741 agatgttgta ccttggagag gaggttaaac tcttgggaga aggcaagatt accgggaaac
     1801 ttgtgggcct gagtgaaaag ggcggtgctt tgatacttac agaggaaggc ataaaggaga
     1861 tactgtccgg cgaatttagc ttgcgacggt cacctcgagc tcggatcatg aggggttacc
     1921 atggcgaccg aggcagccat ccccgcccag cccgctttgc tgaccaacag catatggacg
     1981 tgggcccagc tgccagggcc ccgtacctgc tgggctccag ggaggccttc tccaccgagc
     2041 cccgcttctg tgctccgaga gctggcctgg gacacctttc tccagaagga cccctgagcc
     2101 tgagtgaggg tccatcatca gtaggccctg agggaggccc agggggggtg ggggctgggg
     2161 gaggcagcag taccttcccc aggatgtacc ccggccaggg ccccttcgac acctgtgaag
     2221 actgtgtggg ccacccacag ggcaagggtg ccacccgact gcctcccaca ctcctggacc
     2281 agtttgaaaa gcaattgcca gtccaacaag atggcttcca cacactccca taccagcgag
     2341 gtccagcagg gcctggtcct ggacctggat ctggcgctgc ccccgaggct cgcagcgaga
     2401 gtcctagccg catccggcac ctggttcact cagtgcagaa gctctttgcc aagtcccact
     2461 ctctggaggc acccgggaag cgagactata acgggcccaa ggccgacgga agaggtagct
     2521 ctgggggaga cagctactct ggtccaggct ctggaggcac ccccacctcc caccaccacc
     2581 atcaccacca ccatcaccac caccaccagt cccggcatgg caaacggagc aagagcaagg
     2641 accgcaaggg ggatgggcga caccagacca aggccacagg ctggtggagc tctgatgaca
     2701 atttggacag tgacagtggc tttctgggtg ggagacctcc tggagagcct ggtggcccct
     2761 tctgcctgga tgctccagat gggtcctaca gggacttgag cttcaagggg cgttcgggcg
     2821 ggtcggaagg ccgctgcctt gcctgcactg gcatgtccat gtccctggat ggacagtcag
     2881 tcaagcgaag tgcctggcat accatgatgg tcagccaggg ccgggatgga tacccagggg
     2941 ccgggccagg caaggggctc ctgggtccag agaccaaggc caaagccagg acttatcact
     3001 atttgcaggt gccgcaagat gactgggggg gttaccccac gggtggcaag gatggggaga
     3061 tcccctgccg caggatgcgg agcggcagct acatcaaagc catgggggat gaggagagtg
     3121 gagactcaga tggtagcccc aagacgtctc caaaagcact tgcccgacgc tttgcctccc
     3181 gccgttcctc cagcgtggac acagccagga tcaactgctg tgtcccaccc cggatccatc
     3241 cccggagttc catccctggc tacagccgtt ccctcacaac tggacagcta agcgaagagt
     3301 ttaaccagca gctggaggcc gtgtgcgggt ccgtgtttgg ggagcttgag tcccaggccg
     3361 tggacgccct ggacctgccc ggctgtttcc gcatgcggag ccacagctac ctccgggcca
     3421 tccaggccgg ctgctctcaa gacgacgact gcctacccct ccttgctgcc cctgcctccg
     3481 tctcagggag gcccggctcc tccttcaact tcagaaaggc cccgcccccc atcccgccgg
     3541 gaagccaggc cccaccccgc atctccatca ccgcccagag cagcaccgac tctgcccacg
     3601 agagcttcac cgcagcagag ggcccggccc ggcgctgcag ctccgcggac gggctggacg
     3661 gccccaccat gggcgcacgc accctcgagt tggcaccggt gccaccccga gccagtccca
     3721 agccccccac actcatcatc aagaccatcc ctggccggga ggagctgcgg agcttggcac
     3781 gacagcggaa gtggcggccg tccattgggg tgcaggtgga gacaatctca gactcggaca
     3841 ccgagaacag gagtcgaaga gaattccact ccattggcgt tcaggtggaa gaggacaaga
     3901 ggcgggcgag gttcaagcgc tccaacagcg tgacagccgg tgtgcaggca gacctggagc
     3961 tggagggcct ggccggcctg gccactgtgg ccacagaaga caaggccctg cagttcggac
     4021 gctccttcca gaggcacgcc tctgagcccc agcccggccc tcgggccccc acctactcag
     4081 tcttccgcac ggtccacaca cagggccagt gggcctaccg cgagggctac ccactgccgt
     4141 atgagccgcc ggccaccgat gggtcgcctg gccctacccc tgtcccagcc cccggtcccg
     4201 ggtccggccg ccgtgattcc tggatggagc ggggttcacg tagcctcccc gactcaggcc
     4261 gcacgtcccc ctgcccacgg gatggcgagt ggttcatcaa gatgctgcgg gcagaggtgg
     4321 agaaactgga acactggtgc cagcagatgg agcgtgaggc ggaggactat gagctgcccg
     4381 aggagatctt agagaagatt cgcagtgccg tgggcagtac gcaacttctc ctgtctcaga
     4441 aggtccagca gttcttcagg ctgtgtcagc aaagcttgga ccccactgcg ttccctgtgc
     4501 ccaccttcca ggacctggcc ggtttctggg acctcctgca gctctccatt gaggatgtga
     4561 ccctcaagtt cctggaacta caacaactca aggccaacag ctggaaactc ctggagccga
     4621 aggaggagaa gaaggtccct ccgccgatac caaagaagcc ctcgcggggg cggggcgtgc
     4681 cggtgaagga gcgctccctg gactcggtgg accggcagcg gcaggaagcg cgcaagcggc
     4741 tcctggcggc caagcgcgcc gcctcgttcc gccacagttc ggctaccgag agcgcggaca
     4801 gcatcgagat ctacatcccc gaggcccaga ccaggctgta ggatccaatt cgcttatcga
     4861 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4921 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4981 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5041 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5101 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5161 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     5221 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5281 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5341 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5401 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     5461 gaattctgca gatatctaga gtcgacccgg gcggccgctt cgagcagaca tgataagata
     5521 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     5581 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     5641 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     5701 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     5761 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     5821 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     5881 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta
     5941 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     6001 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     6061 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     6121 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     6181 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     6241 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     6301 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     6361 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6421 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     6481 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6541 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7861 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8101 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     8401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8461 cagcgagtca gtgagcgagg aagcggaag
//
