LOCUS       sequence-478667-        7496 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  to express 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 Jul 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
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     442..461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1412..1421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3160..3748
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3213..3233)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3631..3642)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3875..4269
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   4553..4682
                     /label=AAV2 ITR
     rep_origin      5053..5566
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5140..5159)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5350..5371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6048..6067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6167..6189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6227..6245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6313..6417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6418..6486,6487..7278)
                     /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(6636..6655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7449..7496,1..541)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
       61 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
      121 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      541 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      601 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      661 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg agcgagcgcg
      721 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc
      781 atgctactta tctacgtagc catgctctag actcgagacg cgtaacaaat accatgtcgc
      841 tggtatggag tataggctgt tgctatgaca ggaactcagg ggtcttaact ggcttgagcg
      901 ctgggagggg gcaagcagcc aggccttgtc tgtaagctga agacctggca gtgctgagct
      961 ggtcagcccc caggacctcc ttttgtgccc acgagtgact caccttggca tagacataat
     1021 ggtcaggggt gggcacgcag cctgcttccc gctgtgctcc aggcctcctt cgatgctttc
     1081 cgagaagtct attgagctgg gagcttgtac tgcacccggg gctgacatcc tggcatcctg
     1141 ggataaaagc agcccacggg gctgcccttg ccatatgcct cactggcggc agagaacaag
     1201 gctctattca gcgagtaccc tggggagttc tccccctagc tgggctgggg cccagctcca
     1261 ccccctcagg ctattcaatg ggggtgcttc caggaagtca ggggcagatt tagtccaacc
     1321 cgttcctcca taaaggccct gacatcccag gagccagcag aggcagggca ggttggagcg
     1381 gagacgcatc acctctgcgc gccgcaccgg tgccaccatg gtgagcaagg gcgaggagac
     1441 cacaatgggc gtaatcaagc ccgacatgaa gatcaagctg aagatggagg gcaacgtgaa
     1501 tggccacgcc ttcgtgatcg agggcgaggg cgagggcaag ccctacgacg gcaccaacac
     1561 catcaacctg gaggtgaagg agggagcccc cctgcccttc tcctacgaca ttctgaccac
     1621 cgcgttcgcc tacggcaaca gggccttcac caagtacccc gacgacatcc ccaactactt
     1681 caagcagtcc ttccccgagg gctactcttg ggagcgcacc atgaccttcg aggacaaggg
     1741 catcgtgaag gtgaagtccg acatctccat ggaggaggac tccttcatct acgagataca
     1801 cctcaagggc gagaacttcc cccccaacgg ccccgtgatg cagaagaaga ccaccggctg
     1861 ggacgcctcc accgagagga tgtacgtgcg cgacggcgtg ctgaagggcg acgtcaagca
     1921 caagctgctg ctggagggcg gcggccacca ccgcgttgac ttcaagacca tctacagggc
     1981 caagaaggcg gtgaagctgc ccgactatca ctttgtggac caccgcatcg agatcctgaa
     2041 ccacgacaag gactacaaca aggtgaccgt ttacgagagc gccgtggccc gcatgatgaa
     2101 gcggctggtg tacatcagca agatcagcgg ccacctgagc ctggaagaga tccagcggat
     2161 cggcaaggtg tccatcaaga acaaccagcg ggacaacatc accggcgtgc tgctgtacct
     2221 gcagggcctg ttcttccaga tcctggaagg cgagaacgag aaggtggaca agctgtacaa
     2281 gaaaatcctg gtggacgacc ggcacaccaa catcctgtgc ctgaaaaccg agtacgacat
     2341 caccgaccgg atgttcccca actgggccat gaaaaccatc aacctgaacg agaacagcga
     2401 gctgatgatc cagcccatca agagcctgct gcagaccatc acccagagcc accgggtgct
     2461 ggaaaagtac atgcccgcca gagtgatcta cctgatcaac cagggcatca accccctgac
     2521 cgtggagccc cagctggtcg agaagatcat cttcttcagc gacatcctgg ccttcagcac
     2581 cctgaccgag aagctgcccg tgaacgaggt ggtcatcctg gtcaatcggt acttcagcat
     2641 ctgcacccgg atcatcagcg cctacggcgg cgaagtgacc aagttcatcg gcgactgcgt
     2701 gatggccagc ttcaccaaag agcagggcga cgccgccatc cggaccagcc tggacatcat
     2761 ctccgagctg aagcagctgc ggcaccacgt ggaggccacc aaccccctgc acctgctgta
     2821 caccggcatc ggcctgagct acggccacgt gatcgagggc aacatgggca gcagcctgaa
     2881 gatggaccac accctgctgg gggacgccgt gaacgtggcc gccaggctgg aagccctgac
     2941 aagacagctg ccctacgccc tggcctttac agccggcgtg aagaagtgct gccaggccca
     3001 gtggaccttc atcaacctgg gagcccacca ggtcaagggc aagcaggaag ccatcgaggt
     3061 ctacaccgtg aatgaggccc agaagtacta cgacaccctg cagatcaccc agctgatccg
     3121 gcagaccctg gaaaacgaca agtaagcggc cgcatcgata atcaacctct ggattacaaa
     3181 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3241 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3301 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3361 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3421 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3481 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3541 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3601 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3661 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3721 cggatctccc tttgggccgc ctccccgcat cgatgccgta tacggtaccc ggggatcctc
     3781 tagtagagtc gaggaattca ccccaccagt gcaggctgcc tatcagaaag tggtggctgg
     3841 tgtggctaat gccctggccc acaagtatca ctaagctcgc tttcttgctg tccaatttct
     3901 attaaaggtt cctttgttcc ctaagtccaa ctactaaact gggggatatt atgaagggcc
     3961 ttgagcatct ggattctgcc taataaaaaa catttatttt cattgcaatg atgtatttaa
     4021 attatttctg aatattttac taaaaaggga atgtgggagg tcagtgcatt taaaacataa
     4081 agaaatgaag agctagttca aaccttggga aaatacacta tatcttaaac tccatgaaag
     4141 aaggtgaggc tgcaaacagc taatgcacat tggcaacagc ccctgatgcc tatgccttat
     4201 tcatccctca gaaaaggatt caagtagagg cttgatttgg aggttaaagt tttgctatgc
     4261 tgtattttac attacttatt gttttagctg tcctcatgaa tgtcttttca ctacccattt
     4321 gcttatcctg catctctcag ccttgactcc actcagttct cttgcttaga gataccacct
     4381 ttcccctgaa gtgttccttc catgttttac ggcgagatgg tttctcctcg cctggccact
     4441 cagccttagt tgtctctgtt gtcttataga ggtctggtcg acccccaacg cgttggagct
     4501 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     4561 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4621 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4681 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4741 aatggcgaat ggcgattccg ttgcaatggc tggcggtaat attgttctgg atattaccag
     4801 caaggccgat agtttgagtt cttctactca ggcaagtgat gttattacta atcaaagaag
     4861 tattgcgaca acggttaatt tgcgtgatgg acagactctt ttactcggtg gcctcactga
     4921 ttataaaaac acttctcagg attctggcgt accgttcctg tctaaaatcc ctttaatcgg
     4981 cctcctgttt agctcccgct ctgattctaa cgaggaaagc acgttatacg tgctcgtcaa
     5041 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5101 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5161 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5221 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5281 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5341 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5401 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5461 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttaa atatttgctt
     5521 atacaatctt cctgtttttg gggcttttct gattatcaac cggggtacat atgattgaca
     5581 tgctagtttt acgattaccg ttcatcgatt ctcttgtttg ctccagactc tcaggcaatg
     5641 acctgatagc ctttgtagag acctctcaaa aatagctacc ctctccggca tgaatttatc
     5701 agctagaacg gttgaatatc atattgatgg tgatttgact gtctccggcc tttctcaccc
     5761 gtttgaatct ttacctacac attactcagg cattgcattt aaaatatatg agggttctaa
     5821 aaatttttat ccttgcgttg aaataaaggc ttctcccgca aaagtattac agggtcataa
     5881 tgtttttggt acaaccgatt tagctttatg ctctgaggct ttattgctta attttgctaa
     5941 ttctttgcct tgcctgtatg atttattgga tgttggaatc gcctgatgcg gtattttctc
     6001 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     6061 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6121 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6181 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6241 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6301 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6361 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6421 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6481 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6541 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6601 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6661 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6721 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6781 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6841 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6901 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6961 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7021 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7081 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7141 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7201 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7261 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7321 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7381 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7441 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaa
//
