LOCUS       sequence-348850-        5823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytosolic eGFP in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  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 5823)
  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
FEATURES             Location/Qualifiers
     source          1..5823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    22..610
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(75..95)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(493..504)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    674..795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(685..704)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     739..758
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   957..1086
                     /label=AAV2 ITR
     primer_bind     complement(1105..1122)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1105..1121)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1114..1136)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1263..1718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1350..1369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1560..1581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1744..1848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1849..1917,1918..2709)
                     /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      2880..3468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3369..3388
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    3756..3777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3792..3797,3798..3815,3816..3822)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3830..3846
                     /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     3835..3857
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3854..3870
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3854..3870
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   3908..4037
                     /label=AAV2 ITR
     intron          4888..5020
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5003..5022
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5065..5083
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5104..5106,5107..5109,5110..5823)
                     /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(5104..5106,5107..5109,5110..5820)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5149..5170)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5410..5429)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5757..5778
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggatccaatt cgcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
       61 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
      121 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
      181 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
      241 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
      301 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
      361 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
      421 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
      481 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
      541 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
      601 gcctccccgc atcgataccg gaattctgca gatatctaga gtcgacccgg gcggccgctt
      661 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      721 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
      781 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
      841 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
      901 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
      961 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     1021 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     1081 gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1141 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1201 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1261 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1321 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1381 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1441 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1501 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1561 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1621 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1681 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     1741 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1801 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1861 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1981 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2161 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2221 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2461 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2581 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2701 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     2761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3541 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3601 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3661 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3721 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     3781 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     3841 taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt
     3901 aattaggctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
     3961 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
     4021 catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat ctacgtagcc
     4081 atgctctagg aagatctaac atatcctggt gtggagtagg ggacgctgct ctgacagagg
     4141 ctcgggggcc tgagctggct ctgtgagctg gggaggaggc agacagccag gccttgtctg
     4201 caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc atgcccagtg
     4261 aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg cttcccgccg
     4321 caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc ttgcattgca
     4381 ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta ggggctgccc
     4441 ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt gcccaggaaa
     4501 ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag gagggctgtc
     4561 tgcttcccag aagtccaagg acacaaatgg gtgaggggag agctctcccc atagctgggc
     4621 tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg cacccgggca
     4681 tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag cagagccaga
     4741 gcaggttgga gaggagacgc atcacctccg ctgctcgcaa gctttattgc ggtagtttat
     4801 cacagttaaa ttgctaacgc agtcagtgct tctgacacaa cagtctcgaa cttaagctgc
     4861 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
     4921 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     4981 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat
     5041 tacagctctt aaggctagag tacttaatac gactcactat aggctagcct cgagctcgga
     5101 tccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     5161 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     5221 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     5281 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     5341 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5401 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5461 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     5521 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     5581 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     5641 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5701 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     5761 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5821 taa
//
