LOCUS       sequence-349196-        7579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Glt-1 fused with GFP at the C-terminus in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7579)
  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 7579)
  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-Glt1-eGFP
FEATURES             Location/Qualifiers
     source          1..7579
                     /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"
     regulatory      3207..3216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3213..3215,3216..3218,3219..3932)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3213..3215,3216..3218,3219..3929)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3258..3279)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3519..3538)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3866..3887
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3967..4555
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4020..4040)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4438..4449)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4604..4725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4615..4634)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4669..4688
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4887..5016
                     /label=AAV2 ITR
     primer_bind     complement(5035..5052)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5035..5051)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5044..5066)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5193..5648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5280..5299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5490..5511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5674..5778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5779..5847,5848..6639)
                     /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      6810..7398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7299..7318
                     /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 taggctagcc
     1441 tcgagctcaa gcttcgaatt atggcatcaa cagagggtgc caacaatatg cccaagcagg
     1501 tagaagtgcg catgcatgac agccacctca gctccgatga gccaaagcac cgaaacctgg
     1561 gcatgcgcat gtgcgacaag ctggggaaaa atctcctgct ctcactgact gtgtttggtg
     1621 tcatcctggg agcagtgtgt ggcgggctgc ttcgcttggc atcgcccatc caccctgatg
     1681 tggtcatgtt gatagccttc ccgggggaca tactcatgag gatgctgaag atgctcatcc
     1741 tccctcttat catctccagt ttaatcacag ggttgtcagg cctggatgct aaagccagcg
     1801 gccgcctagg cacgagagct atggtgtatt acatgtccac gaccatcatt gccgccgtgc
     1861 tgggggtcat cctggtgttg gccatccacc caggcaatcc caaactcaag aagcagctag
     1921 ggcccgggaa gaagaacgac gaggtgtcta gcctggatgc cttcctggat ctcattagaa
     1981 atctcttccc ggagaacctg gtgcaagcct gtttccagca gattcagaca gtgacaaaga
     2041 aagttctggt ggcacctcca tctgaggagg ccaataccac caaggcggtc atctccatgt
     2101 tgaatgaaac catgaacgag gcccctgaag aaactaagat cgttatcaag aagggcctgg
     2161 agttcaagga cgggatgaac gtcttaggtc tgatcggatt ctttattgct ttcggcattg
     2221 ccatggggaa gatgggtgaa caggccaagc tgatggtgga gttcttcaac attctgaatg
     2281 agatcgtgat gaagttagtg atcatgatca tgtggtactc ccctctgggt atcgcctgct
     2341 tgatttgtgg gaagatcatc gccatcaagg acttagaagt ggttgctagg cagctgggga
     2401 tgtacatgat caccgtgatc gtgggcctca tcattcacgg gggcatcttt ctccccttga
     2461 tttactttgt agtgaccaga aaaaatccat tctccttttt tgctggcata ttccaagcct
     2521 ggatcactgc tctgggaact gcttccagtg ctggaacttt gcctgttacc ttccgttgct
     2581 tggaagataa tctagggatt gacaagcgtg tgaccagatt cgtcctccca gtcggagcaa
     2641 ccattcacat ggatggcaca gccctttacg aggctgtggc agccatcttc atagcccaaa
     2701 tgaatggggt catcttggat ggaggtcaga ttgtgactgt aagccttaca gccaccctgg
     2761 cgagcattgg tgcagccagt attcccagcg ccgggctggt caccatgctc ctcattctca
     2821 cagctgtggg cctgccaacg gaggatatca gtctgctggt ggcggtggac tggctgctgg
     2881 atagaatgag aacttcagtc aatgtggtgg gcgattcttt tggggctggg attgtctatc
     2941 acctttccaa gtctgagctg gacaccattg actcccaaca ccgaatgcag gaagacatcg
     3001 agatgaccaa gacgcagtcc atttacgacg acaagaacca cagggaaagc aactctaatc
     3061 agtgtgtcta tgccgcacac aactctgtcg taatagatga gtgcaaggta actctggcgg
     3121 ccaatggaaa gtcagctgac tgcagtgttg aggaagaacc ttggaaatct agaggatccc
     3181 gggtggcatc cctggatcca ccggtcgcca ccatggtgag caagggagag gagctgttca
     3241 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3301 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3361 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     3421 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3481 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3541 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3601 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3661 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3721 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3781 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     3841 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3901 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgaccggtat catcaagctt
     3961 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4021 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4081 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4141 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4201 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4261 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4321 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4381 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4441 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4501 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     4561 tacctctaga gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     4621 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4681 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4741 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     4801 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     4861 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4921 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4981 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     5041 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5101 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5161 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     5221 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5281 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5341 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5401 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5461 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5521 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5581 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5641 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5701 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5761 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5821 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5881 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5941 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6001 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6061 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6121 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6181 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6241 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6301 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6361 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6421 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6481 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6541 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6601 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6661 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6721 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6781 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7381 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7441 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7501 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7561 gtgagcgagg aagcggaag
//
