LOCUS       sequence-471027-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to express a version with reduced constitutive activity (thanks to 
            point mutation F198Y) of the optogenetic tool 2pabPAC, which 
            increases cAMP levels when stimulated by blue light, in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  AUTHORS   Vittani M, Herlo R, Wang X, Christensen MDB, Vo CT, Mishima T, Kusk 
            P, Konno A, Hirai H, Tsuboi T, Kitaguchi T, Kjaerby C, Asiminas A, 
            Yokoyama T, Sakamoto M, Nedergaard M, Hirase H
  TITLE     Cerebral blood flow is modulated by astrocytic cAMP elevation 
            independently of IP(3)R2-mediated Ca(2+) signaling in mice.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Jul 8;122(27):e2422069122. doi: 
            10.1073/pnas.2422069122. Epub 2025 Jul 1.
  PUBMED    40591593
REFERENCE   2  (bases 1 to 7496)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2025-07-8"; volume: "122(27):e2422069122. 
            doi"; pages: " 10.1073/pnas.2422069122. Epub 2025 Jul 1"
COMMENT     Sequence Label: pAAV-mGFAP(ABC1D)-2pabPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    820..1408
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(873..893)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1291..1302)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1535..1929
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   2213..2342
                     /label=AAV2 ITR
     rep_origin      2713..3226
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2800..2819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3010..3031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3708..3727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3827..3849
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3887..3905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3973..4077
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4078..4146,4147..4938)
                     /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(4296..4315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5109..5697
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5598..5617
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      6568..6577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccgaccgg atgttcccca actgggccat gaaaaccatc aacctgaacg agaacagcga
       61 gctgatgatc cagcccatca agagcctgct gcagaccatc acccagagcc accgggtgct
      121 ggaaaagtac atgcccgcca gagtgatcta cctgatcaac cagggcatca accccctgac
      181 cgtggagccc cagctggtcg agaagatcat cttcttcagc gacatcctgg ccttcagcac
      241 cctgaccgag aagctgcccg tgaacgaggt ggtcatcctg gtcaatcggt acttcagcat
      301 ctgcacccgg atcatcagcg cctacggcgg cgaagtgacc aagtacatcg gcgactgcgt
      361 gatggccagc ttcaccaaag agcagggcga cgccgccatc cggaccagcc tggacatcat
      421 ctccgagctg aagcagctgc ggcaccacgt ggaggccacc aaccccctgc acctgctgta
      481 caccggcatc ggcctgagct acggccacgt gatcgagggc aacatgggca gcagcctgaa
      541 gatggaccac accctgctgg gggacgccgt gaacgtggcc gccaggctgg aagccctgac
      601 aagacagctg ccctacgccc tggcctttac agccggcgtg aagaagtgct gccaggccca
      661 gtggaccttc atcaacctgg gagcccacca ggtcaagggc aagcaggaag ccatcgaggt
      721 ctacaccgtg aatgaggccc agaagtacta cgacaccctg cagatcaccc agctgatccg
      781 gcagaccctg gaaaacgaca agtaagcggc cgcatcgata atcaacctct ggattacaaa
      841 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
      901 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
      961 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     1021 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     1081 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     1141 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1201 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     1261 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     1321 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     1381 cggatctccc tttgggccgc ctccccgcat cgatgccgta tacggtaccc ggggatcctc
     1441 tagtagagtc gaggaattca ccccaccagt gcaggctgcc tatcagaaag tggtggctgg
     1501 tgtggctaat gccctggccc acaagtatca ctaagctcgc tttcttgctg tccaatttct
     1561 attaaaggtt cctttgttcc ctaagtccaa ctactaaact gggggatatt atgaagggcc
     1621 ttgagcatct ggattctgcc taataaaaaa catttatttt cattgcaatg atgtatttaa
     1681 attatttctg aatattttac taaaaaggga atgtgggagg tcagtgcatt taaaacataa
     1741 agaaatgaag agctagttca aaccttggga aaatacacta tatcttaaac tccatgaaag
     1801 aaggtgaggc tgcaaacagc taatgcacat tggcaacagc ccctgatgcc tatgccttat
     1861 tcatccctca gaaaaggatt caagtagagg cttgatttgg aggttaaagt tttgctatgc
     1921 tgtattttac attacttatt gttttagctg tcctcatgaa tgtcttttca ctacccattt
     1981 gcttatcctg catctctcag ccttgactcc actcagttct cttgcttaga gataccacct
     2041 ttcccctgaa gtgttccttc catgttttac ggcgagatgg tttctcctcg cctggccact
     2101 cagccttagt tgtctctgtt gtcttataga ggtctggtcg acccccaacg cgttggagct
     2161 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     2221 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2281 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2341 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2401 aatggcgaat ggcgattccg ttgcaatggc tggcggtaat attgttctgg atattaccag
     2461 caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta atcaaagaag
     2521 tattgcgaca acggttaatt tgcgtgatgg acagactctt ttactcggtg gcctcactga
     2581 ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc ctttaatcgg
     2641 cctcctgttt agctcccgct ctgattctaa cgaggaaagc acgttatacg tgctcgtcaa
     2701 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2761 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2821 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2881 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2941 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3001 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3061 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3121 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttaa atatttgctt
     3181 atacaatctt cctgtttttg gggcttttct gattatcaac cggggtacat atgattgaca
     3241 tgctagtttt acgattaccg ttcatcgatt ctcttgtttg ctccagactc tcaggcaatg
     3301 acctgatagc ctttgtagag acctctcaaa aatagctacc ctctccggca tgaatttatc
     3361 agctagaacg gttgaatatc atattgatgg tgatttgact gtctccggcc tttctcaccc
     3421 gtttgaatct ttacctacac attactcagg cattgcattt aaaatatatg agggttctaa
     3481 aaatttttat ccttgcgttg aaataaaggc ttctcccgca aaagtattac agggtcataa
     3541 tgtttttggt acaaccgatt tagctttatg ctctgaggct ttattgctta attttgctaa
     3601 ttctttgcct tgcctgtatg atttattgga tgttggaatc gcctgatgcg gtattttctc
     3661 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     3721 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3781 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3841 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3901 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3961 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4021 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4081 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4141 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4201 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4261 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4321 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4381 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4441 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4501 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4561 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4621 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4681 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4741 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4801 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4861 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4921 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4981 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5041 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5101 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5161 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5221 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5281 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5341 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5401 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5461 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5521 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5581 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5641 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5701 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5761 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5821 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagcg agcgcgcaga
     5881 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
     5941 tacttatcta cgtagccatg ctctagactc gagacgcgta acaaatacca tgtcgctggt
     6001 atggagtata ggctgttgct atgacaggaa ctcaggggtc ttaactggct tgagcgctgg
     6061 gagggggcaa gcagccaggc cttgtctgta agctgaagac ctggcagtgc tgagctggtc
     6121 agcccccagg acctcctttt gtgcccacga gtgactcacc ttggcataga cataatggtc
     6181 aggggtgggc acgcagcctg cttcccgctg tgctccaggc ctccttcgat gctttccgag
     6241 aagtctattg agctgggagc ttgtactgca cccggggctg acatcctggc atcctgggat
     6301 aaaagcagcc cacggggctg cccttgccat atgcctcact ggcggcagag aacaaggctc
     6361 tattcagcga gtaccctggg gagttctccc cctagctggg ctggggccca gctccacccc
     6421 ctcaggctat tcaatggggg tgcttccagg aagtcagggg cagatttagt ccaacccgtt
     6481 cctccataaa ggccctgaca tcccaggagc cagcagaggc agggcaggtt ggagcggaga
     6541 cgcatcacct ctgcgcgccg caccggtgcc accatggtga gcaagggcga ggagaccaca
     6601 atgggcgtaa tcaagcccga catgaagatc aagctgaaga tggagggcaa cgtgaatggc
     6661 cacgccttcg tgatcgaggg cgagggcgag ggcaagccct acgacggcac caacaccatc
     6721 aacctggagg tgaaggaggg agcccccctg cccttctcct acgacattct gaccaccgcg
     6781 ttcgcctacg gcaacagggc cttcaccaag taccccgacg acatccccaa ctacttcaag
     6841 cagtccttcc ccgagggcta ctcttgggag cgcaccatga ccttcgagga caagggcatc
     6901 gtgaaggtga agtccgacat ctccatggag gaggactcct tcatctacga gatacacctc
     6961 aagggcgaga acttcccccc caacggcccc gtgatgcaga agaagaccac cggctgggac
     7021 gcctccaccg agaggatgta cgtgcgcgac ggcgtgctga agggcgacgt caagcacaag
     7081 ctgctgctgg agggcggcgg ccaccaccgc gttgacttca agaccatcta cagggccaag
     7141 aaggcggtga agctgcccga ctatcacttt gtggaccacc gcatcgagat cctgaaccac
     7201 gacaaggact acaacaaggt gaccgtttac gagagcgccg tggcccgcat gatgaagcgg
     7261 ctggtgtaca tcagcaagat cagcggccac ctgagcctgg aagagatcca gcggatcggc
     7321 aaggtgtcca tcaagaacaa ccagcgggac aacatcaccg gcgtgctgct gtacctgcag
     7381 ggcctgttct tccagatcct ggaaggcgag aacgagaagg tggacaagct gtacaagaaa
     7441 atcctggtgg acgaccggca caccaacatc ctgtgcctga aaaccgagta cgacat
//
