LOCUS       sequence-371185-        8104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SAPAP3 protein fused with an HA tag at the N-terminus in 
            astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8104)
  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 8104)
  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-3xHA-SAPAP3
FEATURES             Location/Qualifiers
     source          1..8104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    154..175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(190..195,196..213,214..220)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    228..244
                     /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     233..255
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     252..268
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     252..268
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   306..435
                     /label=AAV2 ITR
     intron          1286..1418
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1401..1420
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1463..1481
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1489..1569
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             2233..2262
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2236..2265
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2239..2268
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    4524..5112
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4577..4597)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4995..5006)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5176..5297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5187..5206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5241..5260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5459..5588
                     /label=AAV2 ITR
     primer_bind     complement(5607..5624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5607..5623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5616..5638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5765..6220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5852..5871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6062..6083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6246..6350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6351..6419,6420..7211)
                     /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      7382..7970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7871..7890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
       61 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      121 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      181 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      241 acaatttcac acaggaaaca gctatgacca tgattacgcc agatttaatt aaggccttaa
      301 ttaggctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga
      361 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      421 tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat
      481 gctctaggaa gatctaacat atcctggtgt ggagtagggg acgctgctct gacagaggct
      541 cgggggcctg agctggctct gtgagctggg gaggaggcag acagccaggc cttgtctgca
      601 agcagacctg gcagcattgg gctggccgcc ccccagggcc tcctcttcat gcccagtgaa
      661 tgactcacct tggcacagac acaatgttcg gggtgggcac agtgcctgct tcccgccgca
      721 ccccagcccc cctcaaatgc cttccgagaa gcccattgag cagggggctt gcattgcacc
      781 ccagcctgac agcctggcat cttgggataa aagcagcaca gccccctagg ggctgccctt
      841 gctgtgtggc gccaccggcg gtggagaaca aggctctatt cagcctgtgc ccaggaaagg
      901 ggatcagggg atgcccaggc atggacagtg ggtggcaggg ggggagagga gggctgtctg
      961 cttcccagaa gtccaaggac acaaatgggt gaggggagag ctctccccat agctgggctg
     1021 cggcccaacc ccaccccctc aggctatgcc agggggtgtt gccaggggca cccgggcatc
     1081 gccagtctag cccactcctt cataaagccc tcgcatccca ggagcgagca gagccagagc
     1141 aggttggaga ggagacgcat cacctccgct gctcgcaagc tttattgcgg tagtttatca
     1201 cagttaaatt gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag
     1261 aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga
     1321 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     1381 tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta
     1441 cagctcttaa ggctagagta cttaatacga ctcactatag gctagatgta cccatacgat
     1501 gttccagatt acgcttaccc atacgatgtt ccagattacg cttacccata cgatgttcca
     1561 gattacgcta tgaggggtta ccatggcgac cgaggcagcc atccccgccc agcccgcttt
     1621 gctgaccaac agcatatgga cgtgggccca gctgccaggg ccccgtacct gctgggctcc
     1681 agggaggcct tctccaccga gccccgcttc tgtgctccga gagctggcct gggacacctt
     1741 tctccagaag gacccctgag cctgagtgag ggtccatcat cagtaggccc tgagggaggc
     1801 ccaggggggg tgggggctgg gggaggcagc agtaccttcc ccaggatgta ccccggccag
     1861 ggccccttcg acacctgtga agactgtgtg ggccacccac agggcaaggg tgccacccga
     1921 ctgcctccca cactcctgga ccagtttgaa aagcaattgc cagtccaaca agatggcttc
     1981 cacacactcc cataccagcg aggtccagca gggcctggtc ctggacctgg atctggcgct
     2041 gcccccgagg ctcgcagcga gagtcctagc cgcatccggc acctggttca ctcagtgcag
     2101 aagctctttg ccaagtccca ctctctggag gcacccggga agcgagacta taacgggccc
     2161 aaggccgacg gaagaggtag ctctggggga gacagctact ctggtccagg ctctggaggc
     2221 acccccacct cccaccacca ccatcaccac caccatcacc accaccacca gtcccggcat
     2281 ggcaaacgga gcaagagcaa ggaccgcaag ggggatgggc gacaccagac caaggccaca
     2341 ggctggtgga gctctgatga caatttggac agtgacagtg gctttctggg tgggagacct
     2401 cctggagagc ctggtggccc cttctgcctg gatgctccag atgggtccta cagggacttg
     2461 agcttcaagg ggcgttcggg cgggtcggaa ggccgctgcc ttgcctgcac tggcatgtcc
     2521 atgtccctgg atggacagtc agtcaagcga agtgcctggc ataccatgat ggtcagccag
     2581 ggccgggatg gatacccagg ggccgggcca ggcaaggggc tcctgggtcc agagaccaag
     2641 gccaaagcca ggacttatca ctatttgcag gtgccgcaag atgactgggg gggttacccc
     2701 acgggtggca aggatgggga gatcccctgc cgcaggatgc ggagcggcag ctacatcaaa
     2761 gccatggggg atgaggagag tggagactca gatggtagcc ccaagacgtc tccaaaagca
     2821 cttgcccgac gctttgcctc ccgccgttcc tccagcgtgg acacagccag gatcaactgc
     2881 tgtgtcccac cccggatcca tccccggagt tccatccctg gctacagccg ttccctcaca
     2941 actggacagc taagcgaaga gtttaaccag cagctggagg ccgtgtgcgg gtccgtgttt
     3001 ggggagcttg agtcccaggc cgtggacgcc ctggacctgc ccggctgttt ccgcatgcgg
     3061 agccacagct acctccgggc catccaggcc ggctgctctc aagacgacga ctgcctaccc
     3121 ctccttgctg cccctgcctc cgtctcaggg aggcccggct cctccttcaa cttcagaaag
     3181 gccccgcccc ccatcccgcc gggaagccag gccccacccc gcatctccat caccgcccag
     3241 agcagcaccg actctgccca cgagagcttc accgcagcag agggcccggc ccggcgctgc
     3301 agctccgcgg acgggctgga cggccccacc atgggcgcac gcaccctcga gttggcaccg
     3361 gtgccacccc gagccagtcc caagcccccc acactcatca tcaagaccat ccctggccgg
     3421 gaggagctgc ggagcttggc acgacagcgg aagtggcggc cgtccattgg ggtgcaggtg
     3481 gagacaatct cagactcgga caccgagaac aggagtcgaa gagaattcca ctccattggc
     3541 gttcaggtgg aagaggacaa gaggcgggcg aggttcaagc gctccaacag cgtgacagcc
     3601 ggtgtgcagg cagacctgga gctggagggc ctggccggcc tggccactgt ggccacagaa
     3661 gacaaggccc tgcagttcgg acgctccttc cagaggcacg cctctgagcc ccagcccggc
     3721 cctcgggccc ccacctactc agtcttccgc acggtccaca cacagggcca gtgggcctac
     3781 cgcgagggct acccactgcc gtatgagccg ccggccaccg atgggtcgcc tggccctacc
     3841 cctgtcccag cccccggtcc cgggtccggc cgccgtgatt cctggatgga gcggggttca
     3901 cgtagcctcc ccgactcagg ccgcacgtcc ccctgcccac gggatggcga gtggttcatc
     3961 aagatgctgc gggcagaggt ggagaaactg gaacactggt gccagcagat ggagcgtgag
     4021 gcggaggact atgagctgcc cgaggagatc ttagagaaga ttcgcagtgc cgtgggcagt
     4081 acgcaacttc tcctgtctca gaaggtccag cagttcttca ggctgtgtca gcaaagcttg
     4141 gaccccactg cgttccctgt gcccaccttc caggacctgg ccggtttctg ggacctcctg
     4201 cagctctcca ttgaggatgt gaccctcaag ttcctggaac tacaacaact caaggccaac
     4261 agctggaaac tcctggagcc gaaggaggag aagaaggtcc ctccgccgat accaaagaag
     4321 ccctcgcggg ggcggggcgt gccggtgaag gagcgctccc tggactcggt ggaccggcag
     4381 cggcaggaag cgcgcaagcg gctcctggcg gccaagcgcg ccgcctcgtt ccgccacagt
     4441 tcggctaccg agagcgcgga cagcatcgag atctacatcc ccgaggccca gaccaggctg
     4501 taggatccaa ttcgcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     4561 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4621 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4681 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4741 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4801 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4861 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4921 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     4981 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5041 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5101 ccgcctcccc gcatcgatac cggaattctg cagatatcta gagtcgaccc gggcggccgc
     5161 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     5221 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5281 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     5341 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     5401 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     5461 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     5521 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5581 gcgcgcagcc ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa
     5641 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5701 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5761 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5821 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5881 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5941 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6001 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6061 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6121 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6181 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     6241 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6301 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6361 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6421 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6481 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6541 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6601 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6661 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6721 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6781 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6841 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6901 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6961 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7021 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7081 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7141 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7201 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7261 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7321 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7381 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7441 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7501 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7561 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7621 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7681 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7741 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7801 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7861 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7921 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7981 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8041 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8101 gccg
//
