LOCUS       sequence-422347-        6378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To study calcium signaling in Astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6378)
  AUTHORS   Ollivier M, Soto JS, Linker KE, Moye SL, Jami-Alahmadi Y, Jones AE, 
            Divakaruni AS, Kawaguchi R, Wohlschlegel JA, Khakh BS
  TITLE     Crym-positive striatal astrocytes gate perseverative behaviour.
  JOURNAL   Nature. 2024 Mar;627(8003):358-366. doi: 10.1038/s41586-024-07138-0.
            Epub 2024 Feb 28.
  PUBMED    38418885
REFERENCE   2  (bases 1 to 6378)
  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/s41586-024-07138-0"; journalName: "Nature"; date: 
            "2024-03"; volume: "627"; issue: "8003"; pages: "358-366"
COMMENT     Sequence Label: GfaABC1D-jGCaMP8m
FEATURES             Location/Qualifiers
     source          1..6378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1029..1158
                     /label=AAV2 ITR
     intron          2009..2141
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2124..2143
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        2186..2204
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2234..2251
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     2533..2554
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2660..2681)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2921..2940)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3521..4109
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3574..3594)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3992..4003)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4173..4294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4184..4203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4238..4257
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4456..4585
                     /label=AAV2 ITR
     primer_bind     complement(4604..4621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4604..4620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4613..4635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4762..5217
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4849..4868)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5059..5080
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5243..5347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5348..5416,5417..6208)
                     /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"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccagattta attaaggcct
     1021 taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     1081 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1141 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
     1201 catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc tctgacagag
     1261 gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca ggccttgtct
     1321 gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt catgcccagt
     1381 gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct gcttcccgcc
     1441 gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg cttgcattgc
     1501 accccagcct gacagcctgg catcttggga taaaagcagc acagccccct aggggctgcc
     1561 cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg tgcccaggaa
     1621 aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga ggagggctgt
     1681 ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc catagctggg
     1741 ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg gcacccgggc
     1801 atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga gcagagccag
     1861 agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg cggtagttta
     1921 tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga acttaagctg
     1981 cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg
     2041 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     2101 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa
     2161 ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc tcgagctcgg
     2221 atccaccacc atgcatcatc accatcatca cacgcgtcgc aagaagacct tcaaggaggt
     2281 ggccaccgcc gtgaagatca tcggccgcct gatgggcctg aagatcaacg tctatatcaa
     2341 ggccgacaag cagaagaacg gcatcaaggc gaacttccac atccgccaca acatcgagga
     2401 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
     2461 gctgctgccc gacaaccact acctgagcgt ggagtccaaa ctttcgaaag accccaacga
     2521 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2581 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
     2641 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2701 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2761 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2821 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2881 gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2941 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3001 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca
     3061 actgactgaa gagcagatcg cagaatacaa agaggctttc tccctatttg acaaggacgg
     3121 ggatgggaca ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc
     3181 cacagaagca gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat
     3241 cgacttccct gagttcctga caatgatggc aagaaaaatg aaatacaggg acacggaaga
     3301 agaaattaga gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc
     3361 agagcttcgc cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga
     3421 aatgatcagg gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca
     3481 aatgatgaca gcgaagtaag gatccaattc gcttatcgat aatcaacctc tggattacaa
     3541 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3601 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3661 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3721 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3781 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3841 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3901 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3961 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4021 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4081 tcggatctcc ctttgggccg cctccccgca tcgataccgg aattctgcag atatctagag
     4141 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     4201 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4261 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4321 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4381 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     4441 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4501 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4561 cctcagtgag cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac
     4621 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4681 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4741 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4801 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4861 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4921 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4981 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5041 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5101 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5161 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5221 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5281 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5341 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5401 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5461 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5521 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5581 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5641 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5701 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5761 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5821 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5881 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5941 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6001 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6061 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6121 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6181 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6241 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6301 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6361 aaagatcaaa ggatcttc
//
