LOCUS       sequence-348812-        8468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SAPAP3 protein fused with eGFP at the N-terminus in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8468)
  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 8468)
  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-eGFP-SAPAP3
FEATURES             Location/Qualifiers
     source          1..8468
                     /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             join(1155..1157,1158..1160,1161..1871)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1155..1157,1158..1160,1161..1871)
                     /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(1200..1221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1461..1480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1808..1829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2550..2579
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2553..2582
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2556..2585
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    4841..5429
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4894..4914)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5312..5323)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5493..5614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5504..5523)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5558..5577
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5776..5905
                     /label=AAV2 ITR
     primer_bind     complement(5924..5941)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5924..5940)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5933..5955)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6082..6537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6169..6188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6379..6400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6668..6736,6737..7528)
                     /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(6886..6905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7699..8287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8188..8207
                     /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 taggatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1201 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1261 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1321 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1381 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1441 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1501 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1561 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1621 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1681 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1741 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1801 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1861 agctgtacaa gcctcgagct cggatcatga ggggttacca tggcgaccga ggcagccatc
     1921 cccgcccagc ccgctttgct gaccaacagc atatggacgt gggcccagct gccagggccc
     1981 cgtacctgct gggctccagg gaggccttct ccaccgagcc ccgcttctgt gctccgagag
     2041 ctggcctggg acacctttct ccagaaggac ccctgagcct gagtgagggt ccatcatcag
     2101 taggccctga gggaggccca gggggggtgg gggctggggg aggcagcagt accttcccca
     2161 ggatgtaccc cggccagggc cccttcgaca cctgtgaaga ctgtgtgggc cacccacagg
     2221 gcaagggtgc cacccgactg cctcccacac tcctggacca gtttgaaaag caattgccag
     2281 tccaacaaga tggcttccac acactcccat accagcgagg tccagcaggg cctggtcctg
     2341 gacctggatc tggcgctgcc cccgaggctc gcagcgagag tcctagccgc atccggcacc
     2401 tggttcactc agtgcagaag ctctttgcca agtcccactc tctggaggca cccgggaagc
     2461 gagactataa cgggcccaag gccgacggaa gaggtagctc tgggggagac agctactctg
     2521 gtccaggctc tggaggcacc cccacctccc accaccacca tcaccaccac catcaccacc
     2581 accaccagtc ccggcatggc aaacggagca agagcaagga ccgcaagggg gatgggcgac
     2641 accagaccaa ggccacaggc tggtggagct ctgatgacaa tttggacagt gacagtggct
     2701 ttctgggtgg gagacctcct ggagagcctg gtggcccctt ctgcctggat gctccagatg
     2761 ggtcctacag ggacttgagc ttcaaggggc gttcgggcgg gtcggaaggc cgctgccttg
     2821 cctgcactgg catgtccatg tccctggatg gacagtcagt caagcgaagt gcctggcata
     2881 ccatgatggt cagccagggc cgggatggat acccaggggc cgggccaggc aaggggctcc
     2941 tgggtccaga gaccaaggcc aaagccagga cttatcacta tttgcaggtg ccgcaagatg
     3001 actggggggg ttaccccacg ggtggcaagg atggggagat cccctgccgc aggatgcgga
     3061 gcggcagcta catcaaagcc atgggggatg aggagagtgg agactcagat ggtagcccca
     3121 agacgtctcc aaaagcactt gcccgacgct ttgcctcccg ccgttcctcc agcgtggaca
     3181 cagccaggat caactgctgt gtcccacccc ggatccatcc ccggagttcc atccctggct
     3241 acagccgttc cctcacaact ggacagctaa gcgaagagtt taaccagcag ctggaggccg
     3301 tgtgcgggtc cgtgtttggg gagcttgagt cccaggccgt ggacgccctg gacctgcccg
     3361 gctgtttccg catgcggagc cacagctacc tccgggccat ccaggccggc tgctctcaag
     3421 acgacgactg cctacccctc cttgctgccc ctgcctccgt ctcagggagg cccggctcct
     3481 ccttcaactt cagaaaggcc ccgcccccca tcccgccggg aagccaggcc ccaccccgca
     3541 tctccatcac cgcccagagc agcaccgact ctgcccacga gagcttcacc gcagcagagg
     3601 gcccggcccg gcgctgcagc tccgcggacg ggctggacgg ccccaccatg ggcgcacgca
     3661 ccctcgagtt ggcaccggtg ccaccccgag ccagtcccaa gccccccaca ctcatcatca
     3721 agaccatccc tggccgggag gagctgcgga gcttggcacg acagcggaag tggcggccgt
     3781 ccattggggt gcaggtggag acaatctcag actcggacac cgagaacagg agtcgaagag
     3841 aattccactc cattggcgtt caggtggaag aggacaagag gcgggcgagg ttcaagcgct
     3901 ccaacagcgt gacagccggt gtgcaggcag acctggagct ggagggcctg gccggcctgg
     3961 ccactgtggc cacagaagac aaggccctgc agttcggacg ctccttccag aggcacgcct
     4021 ctgagcccca gcccggccct cgggccccca cctactcagt cttccgcacg gtccacacac
     4081 agggccagtg ggcctaccgc gagggctacc cactgccgta tgagccgccg gccaccgatg
     4141 ggtcgcctgg ccctacccct gtcccagccc ccggtcccgg gtccggccgc cgtgattcct
     4201 ggatggagcg gggttcacgt agcctccccg actcaggccg cacgtccccc tgcccacggg
     4261 atggcgagtg gttcatcaag atgctgcggg cagaggtgga gaaactggaa cactggtgcc
     4321 agcagatgga gcgtgaggcg gaggactatg agctgcccga ggagatctta gagaagattc
     4381 gcagtgccgt gggcagtacg caacttctcc tgtctcagaa ggtccagcag ttcttcaggc
     4441 tgtgtcagca aagcttggac cccactgcgt tccctgtgcc caccttccag gacctggccg
     4501 gtttctggga cctcctgcag ctctccattg aggatgtgac cctcaagttc ctggaactac
     4561 aacaactcaa ggccaacagc tggaaactcc tggagccgaa ggaggagaag aaggtccctc
     4621 cgccgatacc aaagaagccc tcgcgggggc ggggcgtgcc ggtgaaggag cgctccctgg
     4681 actcggtgga ccggcagcgg caggaagcgc gcaagcggct cctggcggcc aagcgcgccg
     4741 cctcgttccg ccacagttcg gctaccgaga gcgcggacag catcgagatc tacatccccg
     4801 aggcccagac caggctgtag gatccaattc gcttatcgat aatcaacctc tggattacaa
     4861 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4921 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4981 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5041 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5101 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     5161 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     5221 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     5281 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     5341 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     5401 tcggatctcc ctttgggccg cctccccgca tcgataccgg aattctgcag atatctagag
     5461 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     5521 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     5581 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     5641 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     5701 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     5761 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     5821 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     5881 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac
     5941 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     6001 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     6061 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6421 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6481 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8101 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     8341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     8401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     8461 agcggaag
//
