LOCUS       sequence-348831-        7634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ezrin fused with eGFP at the C-terminus in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7634)
  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 7634)
  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-Ezr-eGFP
FEATURES             Location/Qualifiers
     source          1..7634
                     /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      3262..3271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3268..3270,3271..3273,3274..3987)
                     /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(3268..3270,3271..3273,3274..3984)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3313..3334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3574..3593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3921..3942
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4022..4610
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4075..4095)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4493..4504)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4659..4780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4670..4689)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4724..4743
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4942..5071
                     /label=AAV2 ITR
     primer_bind     complement(5090..5107)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5090..5106)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5099..5121)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5248..5703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5335..5354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5545..5566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5729..5833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5834..5902,5903..6694)
                     /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      6865..7453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7354..7373
                     /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 caccaccatg ccgaaaccaa tcaatgtccg agttaccacc
     1501 atggatgcag agctggagtt tgcaatccag ccaaatacaa ctggaaaaca gctttttgat
     1561 caggtggtaa agactatcgg cctccgggaa gtgtggtact ttggcctcca ctatgtggat
     1621 aataaaggat ttcctacctg gctgaagctg gataagaagg tgtctgccca ggaggtcagg
     1681 aaggagaatc ccctccagtt caagttccgg gccaagttct accctgaaga tgtggctgag
     1741 gagctcatcc aggacatcac ccagaaactt ttcttcctcc aagtgaagga aggaatcctt
     1801 agcgatgaga tctactgccc ccctgagact gccgtgctct tggggtccta cgctgtgcag
     1861 gccaagtttg gggactacaa caaagaagtg cacaagtctg ggtacctcag ctctgagcgg
     1921 ctgatccctc aaagagtgat ggaccagcac aaacttacca gggaccagtg ggaggaccgg
     1981 atccaggtgt ggcatgcgga acaccgtggg atgctcaaag ataatgctat gttggaatac
     2041 ctgaagattg ctcaggacct ggaaatgtat ggaatcaact atttcgagat aaaaaacaag
     2101 aaaggaacag acctttggct tggagttgat gcccttggac tgaatattta tgagaaagat
     2161 gataagttaa ccccaaagat tggctttcct tggagtgaaa tcaggaacat ctctttcaat
     2221 gacaaaaagt ttgtcattaa acccatcgac aagaaggcac ctgactttgt gttttatgcc
     2281 ccacgtctga gaatcaacaa gcggatcctg cagctctgca tgggcaacca tgagttgtat
     2341 atgcgccgca ggaagcctga caccatcgag gtgcagcaga tgaaggccca ggcccgggag
     2401 gagaagcatc agaagcagct ggagcggcaa cagctggaaa cagagaagaa aaggagagaa
     2461 accgtggaga gagagaaaga gcagatgatg cgcgagaagg aggagttgat gctgcggctg
     2521 caggactatg aggagaagac aaagaaggca gagagagagc tctcggagca gattcagagg
     2581 gccctgcagc tggaggagga gaggaagcgg gcacaggagg aggccgagcg cctagaggct
     2641 gaccgtatgg ctgcactgcg ggctaaggag gagctggaga gacaggcggt ggatcagata
     2701 aagagccagg agcagctggc tgcggagctt gcagaataca ctgccaagat tgccctcctg
     2761 gaagaggcgc ggaggcgcaa ggaggatgaa gttgaagagt ggcagcacag ggccaaagaa
     2821 gcccaggatg acctggtgaa gaccaaggag gagctgcacc tggtgatgac agcacccccg
     2881 cccccaccac cccccgtgta cgagccggtg agctaccatg tccaggagag cttgcaggat
     2941 gagggcgcag agcccacggg ctacagcgcg gagctgtcta gtgagggcat ccgggatgac
     3001 cgcaatgagg agaagcgcat cactgaggca gagaagaacg agcgtgtgca gcggcagctg
     3061 ctgacgctga gcagcgagct gtcccaggcc cgagatgaga ataagaggac ccacaatgac
     3121 atcatccaca acgagaacat gaggcaaggc cgggacaagt acaagacgct gcggcagatc
     3181 cggcagggca acaccaagca gcgcatcgac gagttcgagg ccctgtcgtc gacggtaccg
     3241 cgggcccggg atccaccggt cgccaccatg gtgagcaagg gagaggagct gttcaccggg
     3301 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3361 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     3421 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     3481 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3541 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3601 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3661 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     3721 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     3781 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3841 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     3901 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3961 ctcggcatgg acgagctgta caagtaaagc ggccgcgacc ggtatcatca agcttatcga
     4021 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4081 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4141 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4201 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4261 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4321 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4381 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     4441 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4501 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4561 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgatacct
     4621 ctagagtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4681 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4741 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4801 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     4861 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     4921 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     4981 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     5041 gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg
     5101 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5161 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5221 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     5281 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5341 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5401 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5461 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5521 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5581 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5641 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5701 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5761 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5821 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5881 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5941 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6001 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6061 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6121 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6181 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6241 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6301 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6361 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6421 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6481 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6541 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6601 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6661 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6721 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6781 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6841 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6901 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6961 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7021 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7081 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7141 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7201 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7261 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7321 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7381 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7441 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7501 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7561 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7621 cgaggaagcg gaag
//
