LOCUS       sequence-451914-        6974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to express the red cAMP indicator in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6974)
  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 6974)
  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-Lck-PinkFlamindo
FEATURES             Location/Qualifiers
     source          1..6974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1262..1281
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2036..2624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2089..2109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2507..2518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2751..3145
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   3429..3558
                     /label=AAV2 ITR
     rep_origin      3929..4442
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4016..4035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4226..4247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4924..4943)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5043..5065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5103..5121)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5189..5293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5294..5362,5363..6154)
                     /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(5512..5531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6325..6913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6814..6833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
       61 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag cgagcgcgca
      121 gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat
      181 gctacttatc tacgtagcca tgctctagac tcgagacgcg taacaaatac catgtcgctg
      241 gtatggagta taggctgttg ctatgacagg aactcagggg tcttaactgg cttgagcgct
      301 gggagggggc aagcagccag gccttgtctg taagctgaag acctggcagt gctgagctgg
      361 tcagccccca ggacctcctt ttgtgcccac gagtgactca ccttggcata gacataatgg
      421 tcaggggtgg gcacgcagcc tgcttcccgc tgtgctccag gcctccttcg atgctttccg
      481 agaagtctat tgagctggga gcttgtactg cacccggggc tgacatcctg gcatcctggg
      541 ataaaagcag cccacggggc tgcccttgcc atatgcctca ctggcggcag agaacaaggc
      601 tctattcagc gagtaccctg gggagttctc cccctagctg ggctggggcc cagctccacc
      661 ccctcaggct attcaatggg ggtgcttcca ggaagtcagg ggcagattta gtccaacccg
      721 ttcctccata aaggccctga catcccagga gccagcagag gcagggcagg ttggagcgga
      781 gacgcatcac ctctgcgcgc cgcaccggtg ccaccatggg ctgcgtgtgc agcagcaacc
      841 ccgagatgct ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
      901 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
      961 gcgagggccg cccctacgag gcctttcaga ccgctaagct gaaggtgacc aagggtggcc
     1021 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctaca
     1081 ttaagcaccc agccgacatc cccgactact tcaagctgtc cttccccgag ggcttcaggt
     1141 gggagcgcgt gatgaacttc gaggacggcg gcattattca cgttaaccag gactcctccc
     1201 tgcaggacgg cgtattcatc tacaaggtga agctgcgcgg caccaacttc ccccccgacg
     1261 gccccgtaat gcagaagaag accatgggct gggagcagtg gcgcgatcct cctctcctgt
     1321 cccagcgagg gcctgatgcc ctactcaccg tggcgctccg gaagccccca ggtcagcgta
     1381 cggatgaaga actggacctg atctttgaag agctgctgca tatcaaggcg gtggcacacc
     1441 tttctaactc ggtgaagcgg gaactagctg ctgttctgct ctttgaacca cacagcaagg
     1501 caggaactgt gttgttcagc cagggggaca agggtacctc gtggtacatt atctggaagg
     1561 gatctgtcaa tgtggtgacc catggcaagg ggctggtgac cacgttgcac gagggagatg
     1621 actttggaca gctggctctg gtgaacgacg cacctcgggc agccaccatc atccttcgag
     1681 aaaataactg tcactttctg cgtgtggaca agcaggactt caaccgcatc atcaaggatg
     1741 tggaatttaa agtggagact gttgatgagc ggatgtaccc cgaggacggc gccctgaaga
     1801 gcgagatcaa gaaggggctg aggctgaagg acggcggcca ctacgccgcc gaggtcaaga
     1861 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc gacatcaagt
     1921 tggacatcgt gtcccacaac gaggactaca ccatcgtgga acagtgcgaa cgcgccgagg
     1981 gccgccactc caccggcggc atggacgagc tgtacaagta agcggccgca tcgataatca
     2041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     2101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     2161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     2221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     2281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     2341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     2401 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     2461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     2521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     2581 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat gccgtatacg
     2641 gtacccgggg atcctctagt agagtcgagg aattcacccc accagtgcag gctgcctatc
     2701 agaaagtggt ggctggtgtg gctaatgccc tggcccacaa gtatcactaa gctcgctttc
     2761 ttgctgtcca atttctatta aaggttcctt tgttccctaa gtccaactac taaactgggg
     2821 gatattatga agggccttga gcatctggat tctgcctaat aaaaaacatt tattttcatt
     2881 gcaatgatgt atttaaatta tttctgaata ttttactaaa aagggaatgt gggaggtcag
     2941 tgcatttaaa acataaagaa atgaagagct agttcaaacc ttgggaaaat acactatatc
     3001 ttaaactcca tgaaagaagg tgaggctgca aacagctaat gcacattggc aacagcccct
     3061 gatgcctatg ccttattcat ccctcagaaa aggattcaag tagaggcttg atttggaggt
     3121 taaagttttg ctatgctgta ttttacatta cttattgttt tagctgtcct catgaatgtc
     3181 ttttcactac ccatttgctt atcctgcatc tctcagcctt gactccactc agttctcttg
     3241 cttagagata ccacctttcc cctgaagtgt tccttccatg ttttacggcg agatggtttc
     3301 tcctcgcctg gccactcagc cttagttgtc tctgttgtct tatagaggtc tggtcgaccc
     3361 ccaacgcgtt ggagctctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     3421 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3481 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3541 gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3601 acagttgcgc agcctgaatg gcgaatggcg attccgttgc aatggctggc ggtaatattg
     3661 ttctggatat taccagcaag gccgatagtt tgagttcttc tactcaggca agtgatgtta
     3721 ttactaatca aagaagtatt gcgacaacgg ttaatttgcg tgatggacag actcttttac
     3781 tcggtggcct cactgattat aaaaacactt ctcaggattc tggcgtaccg ttcctgtcta
     3841 aaatcccttt aatcggcctc ctgtttagct cccgctctga ttctaacgag gaaagcacgt
     3901 tatacgtgct cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3961 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4021 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4081 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4141 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4201 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4261 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4321 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     4381 atttaaatat ttgcttatac aatcttcctg tttttggggc ttttctgatt atcaaccggg
     4441 gtacatatga ttgacatgct agttttacga ttaccgttca tcgattctct tgtttgctcc
     4501 agactctcag gcaatgacct gatagccttt gtagagacct ctcaaaaata gctaccctct
     4561 ccggcatgaa tttatcagct agaacggttg aatatcatat tgatggtgat ttgactgtct
     4621 ccggcctttc tcacccgttt gaatctttac ctacacatta ctcaggcatt gcatttaaaa
     4681 tatatgaggg ttctaaaaat ttttatcctt gcgttgaaat aaaggcttct cccgcaaaag
     4741 tattacaggg tcataatgtt tttggtacaa ccgatttagc tttatgctct gaggctttat
     4801 tgcttaattt tgctaattct ttgccttgcc tgtatgattt attggatgtt ggaatcgcct
     4861 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     4921 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4981 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5041 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5101 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5161 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5221 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5281 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5341 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5401 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5461 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5521 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5581 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5641 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5701 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5761 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5821 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5881 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5941 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6001 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6061 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6121 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6181 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6241 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6961 ttttgctcac atgt
//
