LOCUS       sequence-348847-        8748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SAPAP3 protein fused with eGFP at the N-terminus in 
            astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8748)
  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 8748)
  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-eGFP-SAPAP3
FEATURES             Location/Qualifiers
     source          1..8748
                     /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             join(1435..1437,1438..1440,1441..2151)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1435..1437,1438..1440,1441..2151)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1480..1501)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1741..1760)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2088..2109
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2830..2859
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2833..2862
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2836..2865
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    5121..5709
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5174..5194)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5592..5603)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5773..5894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5784..5803)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5838..5857
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6056..6185
                     /label=AAV2 ITR
     primer_bind     complement(6204..6221)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6204..6220)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6213..6235)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6362..6817
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6449..6468)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6659..6680
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6843..6947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6948..7016,7017..7808)
                     /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      7979..8567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8468..8487
                     /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 taggatggtg
     1441 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1501 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1561 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1621 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1681 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1741 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1801 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1861 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1921 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1981 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2041 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2101 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gcctcgagct
     2161 cggatcatga ggggttacca tggcgaccga ggcagccatc cccgcccagc ccgctttgct
     2221 gaccaacagc atatggacgt gggcccagct gccagggccc cgtacctgct gggctccagg
     2281 gaggccttct ccaccgagcc ccgcttctgt gctccgagag ctggcctggg acacctttct
     2341 ccagaaggac ccctgagcct gagtgagggt ccatcatcag taggccctga gggaggccca
     2401 gggggggtgg gggctggggg aggcagcagt accttcccca ggatgtaccc cggccagggc
     2461 cccttcgaca cctgtgaaga ctgtgtgggc cacccacagg gcaagggtgc cacccgactg
     2521 cctcccacac tcctggacca gtttgaaaag caattgccag tccaacaaga tggcttccac
     2581 acactcccat accagcgagg tccagcaggg cctggtcctg gacctggatc tggcgctgcc
     2641 cccgaggctc gcagcgagag tcctagccgc atccggcacc tggttcactc agtgcagaag
     2701 ctctttgcca agtcccactc tctggaggca cccgggaagc gagactataa cgggcccaag
     2761 gccgacggaa gaggtagctc tgggggagac agctactctg gtccaggctc tggaggcacc
     2821 cccacctccc accaccacca tcaccaccac catcaccacc accaccagtc ccggcatggc
     2881 aaacggagca agagcaagga ccgcaagggg gatgggcgac accagaccaa ggccacaggc
     2941 tggtggagct ctgatgacaa tttggacagt gacagtggct ttctgggtgg gagacctcct
     3001 ggagagcctg gtggcccctt ctgcctggat gctccagatg ggtcctacag ggacttgagc
     3061 ttcaaggggc gttcgggcgg gtcggaaggc cgctgccttg cctgcactgg catgtccatg
     3121 tccctggatg gacagtcagt caagcgaagt gcctggcata ccatgatggt cagccagggc
     3181 cgggatggat acccaggggc cgggccaggc aaggggctcc tgggtccaga gaccaaggcc
     3241 aaagccagga cttatcacta tttgcaggtg ccgcaagatg actggggggg ttaccccacg
     3301 ggtggcaagg atggggagat cccctgccgc aggatgcgga gcggcagcta catcaaagcc
     3361 atgggggatg aggagagtgg agactcagat ggtagcccca agacgtctcc aaaagcactt
     3421 gcccgacgct ttgcctcccg ccgttcctcc agcgtggaca cagccaggat caactgctgt
     3481 gtcccacccc ggatccatcc ccggagttcc atccctggct acagccgttc cctcacaact
     3541 ggacagctaa gcgaagagtt taaccagcag ctggaggccg tgtgcgggtc cgtgtttggg
     3601 gagcttgagt cccaggccgt ggacgccctg gacctgcccg gctgtttccg catgcggagc
     3661 cacagctacc tccgggccat ccaggccggc tgctctcaag acgacgactg cctacccctc
     3721 cttgctgccc ctgcctccgt ctcagggagg cccggctcct ccttcaactt cagaaaggcc
     3781 ccgcccccca tcccgccggg aagccaggcc ccaccccgca tctccatcac cgcccagagc
     3841 agcaccgact ctgcccacga gagcttcacc gcagcagagg gcccggcccg gcgctgcagc
     3901 tccgcggacg ggctggacgg ccccaccatg ggcgcacgca ccctcgagtt ggcaccggtg
     3961 ccaccccgag ccagtcccaa gccccccaca ctcatcatca agaccatccc tggccgggag
     4021 gagctgcgga gcttggcacg acagcggaag tggcggccgt ccattggggt gcaggtggag
     4081 acaatctcag actcggacac cgagaacagg agtcgaagag aattccactc cattggcgtt
     4141 caggtggaag aggacaagag gcgggcgagg ttcaagcgct ccaacagcgt gacagccggt
     4201 gtgcaggcag acctggagct ggagggcctg gccggcctgg ccactgtggc cacagaagac
     4261 aaggccctgc agttcggacg ctccttccag aggcacgcct ctgagcccca gcccggccct
     4321 cgggccccca cctactcagt cttccgcacg gtccacacac agggccagtg ggcctaccgc
     4381 gagggctacc cactgccgta tgagccgccg gccaccgatg ggtcgcctgg ccctacccct
     4441 gtcccagccc ccggtcccgg gtccggccgc cgtgattcct ggatggagcg gggttcacgt
     4501 agcctccccg actcaggccg cacgtccccc tgcccacggg atggcgagtg gttcatcaag
     4561 atgctgcggg cagaggtgga gaaactggaa cactggtgcc agcagatgga gcgtgaggcg
     4621 gaggactatg agctgcccga ggagatctta gagaagattc gcagtgccgt gggcagtacg
     4681 caacttctcc tgtctcagaa ggtccagcag ttcttcaggc tgtgtcagca aagcttggac
     4741 cccactgcgt tccctgtgcc caccttccag gacctggccg gtttctggga cctcctgcag
     4801 ctctccattg aggatgtgac cctcaagttc ctggaactac aacaactcaa ggccaacagc
     4861 tggaaactcc tggagccgaa ggaggagaag aaggtccctc cgccgatacc aaagaagccc
     4921 tcgcgggggc ggggcgtgcc ggtgaaggag cgctccctgg actcggtgga ccggcagcgg
     4981 caggaagcgc gcaagcggct cctggcggcc aagcgcgccg cctcgttccg ccacagttcg
     5041 gctaccgaga gcgcggacag catcgagatc tacatccccg aggcccagac caggctgtag
     5101 gatccaattc gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     5161 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5221 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5281 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     5341 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     5401 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     5461 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     5521 cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg acgtccttct
     5581 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     5641 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     5701 cctccccgca tcgataccgg aattctgcag atatctagag tcgacccggg cggccgcttc
     5761 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     5821 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     5881 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga
     5941 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     6001 ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa
     6061 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     6121 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     6181 cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6241 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6301 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6361 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6421 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6481 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6541 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6601 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6661 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6721 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6781 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     6841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8641 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8701 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
