LOCUS       sequence-441248-        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing GFAP-GeNL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  AUTHORS   Beinlich FRM, Asiminas A, Untiet V, Bojarowska Z, Pla V, Sigurdsson 
            B, Timmel V, Gehrig L, Graber MH, Hirase H, Nedergaard M
  TITLE     Oxygen imaging of hypoxic pockets in the mouse cerebral cortex.
  JOURNAL   Science. 2024 Mar 29;383(6690):1471-1478. doi: 
            10.1126/science.adn1011. Epub 2024 Mar 28.
  PUBMED    38547288
REFERENCE   2  (bases 1 to 7443)
  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: 
            "10.1126/science.adn1011"; journalName: "Science"; date: 
            "2024-03-29- 29"; volume: "383"; issue: "6690"; pages: "1471-1478"
COMMENT     Sequence Label: pssAAV-2-hGFAP-GeNL-WPRE-bGHp(A)
FEATURES             Location/Qualifiers
     source          1..7443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   191..331
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   191..320
                     /label=AAV2 ITR
     rep_origin      406..861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(493..512)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     703..724
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(878..897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     997..1019
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1057..1075)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1143..1247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1248..1316,1317..2108)
                     /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      2279..2867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2768..2787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2929..3058
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             6607..6648
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    6666..7254
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6719..6739)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7137..7148)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7279..7296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7319..7336)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(7325..7443,1..106)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
       61 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
      121 agaaccagat cctctcttaa ggtagcatcg agatttaaat tagggataac agggtaatgg
      181 cgcgggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
      241 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
      301 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
      361 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
      421 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      481 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      541 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      601 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
      661 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      721 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
      781 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      841 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
      901 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      961 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     1021 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     1081 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1141 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1201 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1261 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1321 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1381 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1441 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1501 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1561 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1621 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1681 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1741 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1801 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1861 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1921 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1981 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2041 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2101 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2161 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2221 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2281 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2341 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2401 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2461 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2521 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2581 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2641 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2701 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2761 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2821 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2881 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     2941 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     3001 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     3061 ggcaattcag tcgataacta taacggtcct aaggtagcga tttaaatacg cgctctctta
     3121 aggtagcccc gggacgcgtc aattgtccca cctccctctc tgtgctggga ctcacagagg
     3181 gagacctcag gaggcagtct gtccatcaca tgtccaaatg cagagcatac cctgggctgg
     3241 gcgcagtggc gcacaactgt aattccagca ctttgggagg ctgatgtgga aggatcactt
     3301 gagcccagaa gttctagacc agcctgggca acatggcaag accctatctc tacaaaaaaa
     3361 gttaaaaaat cagccacgtg tggtgacaca cacctgtagt cccagctatt caggaggctg
     3421 aggtgagggg atcacttaag gctgggaggt tgaggctgca gtgagtcgtg gttgcgccac
     3481 tgcactccag cctgggcaac agtgagaccc tgtctcaaaa gacaaaaaaa aaaaaaaaaa
     3541 aaaaaaaaaa gaacatatcc tggtgtggag taggggacgc tgctctgaca gaggctcggg
     3601 ggcctgagct ggctctgtga gctggggagg aggcagacag ccaggccttg tctgcaagca
     3661 gacctggcag cattgggctg gccgcccccc agggcctcct cttcatgccc agtgaatgac
     3721 tcaccttggc acagacacaa tgttcggggt gggcacagtg cctgcttccc gccgcacccc
     3781 agcccccctc aaatgccttc cgagaagccc attgagtagg gggcttgcat tgcaccccag
     3841 cctgacagcc tggcatcttg ggataaaagc agcacagccc cctaggggct gcccttgctg
     3901 tgtggcgcca ccggcggtgg agaacaaggc tctattcagc ctgtgcccag gaaaggggat
     3961 caggggatgc ccaggcatgg acagtgggtg gcaggggggg agaggagggc tgtctgcttc
     4021 ccagaagtcc aaggacacaa atgggtgagg ggactgggca gggttctgac cctgtgggac
     4081 cagagtggag ggcgtagatg gacctgaagt ctccagggac aacagggccc aggtctcagg
     4141 ctcctagttg ggcccagtgg ctccagcgtt tccaaaccca tccatcccca gaggttcttc
     4201 ccatctctcc aggctgatgt gtgggaactc gaggaaataa atctccagtg ggagacggag
     4261 gggtggccag ggaaacgggg cgctgcagga ataaagacga gccagcacag ccagctcatg
     4321 cgtaacggct ttgtggagct gtcaaggcct ggtctctggg agagaggcac agggaggcca
     4381 gacaaggaag gggtgacctg gagggacaga tccaggggct aaagtcctga taaggcaaga
     4441 gagtgccggc cccctcttgc cctatcagga cctccactgc cacatagagg ccatgattga
     4501 cccttagaca aagggctggt gtccaatccc agcccccagc cccagaactc cagggaatga
     4561 atgggcagag agcaggaatg tgggacatct gtgttcaagg gaaggactcc aggagtctgc
     4621 tgggaatgag gcctagtagg aaatgaggtg gcccttgagg gtacagaaca ggttcattct
     4681 tcgccaaatt cccagcacct tgcaggcact tacagctgag tgagataatg cctgggttat
     4741 gaaatcaaaa agttggaaag caggtcagag gtcatctggt acagcccttc cttccctttt
     4801 tttttttttt ttttttttgt gagacaaggt ctctctctgt tgcccaggct ggagtggcgc
     4861 aaacacagct cactgcagcc tcaacctact gggctcaagc aatcctccag cctcagcctc
     4921 ccaaagtgct gggattacaa gcatgagcca ccccactcag ccctttcctt cctttttaat
     4981 tgatgcataa taattgtaag tattcatcat ggtccaacca accctttctt gacccacctt
     5041 cctagagaga gggtcctctt gattcagcgg tcagggcccc agacccatgg tctggctcca
     5101 ggtaccacct gcctcatgca ggagttggcg tgcccaggaa gctctgcctc tgggcacagt
     5161 gacctcagtg gggtgagggg agctctcccc atagctgggc tgcggcccaa ccccaccccc
     5221 tcaggctatg ccagggggtg ttgccagggg cacccgggca tcgccagtct agcccactcc
     5281 ttcataaagc cctcgcatcc caggagcgag cagagccaga gcagctagcg tttaaactta
     5341 agcttggtac cgagctcgga tccgatggtg tccaagggcg aagaggacaa catggccagc
     5401 ctgcctgcca cccacgagct gcacatcttc ggcagcatca acggcgtgga cttcgacatg
     5461 gtgggacagg gcaccggcaa ccccaacgac ggctacgagg aactgaacct gaagtccaca
     5521 aagggcgacc tgcagttcag cccctggatt ctggtgcccc acatcggcta cggcttccac
     5581 cagtacctgc cctaccccga cggcatgagc cctttccagg ccgctatggt ggatggcagc
     5641 ggctaccagg tgcaccggac catgcagttt gaggacggcg ccagcctgac cgtgaactac
     5701 cggtacacat acgagggcag ccacatcaag ggcgaggccc aagtgaaggg cacaggcttt
     5761 ccagccgacg gccccgtgat gaccaatagc ctgacagccg ccgactggtg cagaagcaag
     5821 aaaacctacc ccaatgacaa gaccatcatc agcaccttca agtggtccta caccaccggc
     5881 aatggcaagc ggtacagaag caccgcccgg accacctaca ccttcgccaa acctatggcc
     5941 gccaactacc tgaagaacca gcctatgtac gtgttccgca agaccgagct gaagcactcc
     6001 aagacagaac tgaacttcaa agagtggcag aaagccttca ccgggtttga agatttcgtt
     6061 ggggactggc gacagacagc cggctacaac ctggaccaag tccttgaaca gggaggtgtg
     6121 tccagtttgt ttcagaatct cggggtgtcc gtaactccga tccaaaggat tgtcctgagc
     6181 ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcggc
     6241 gaccaaatgg gccagatcga aaaaattttt aaggtggtgt accctgtgga tgatcatcac
     6301 tttaaggtga tcctgcacta tggcacactg gtaatcgacg gggttacgcc gaacatgatc
     6361 gactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta
     6421 acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa ccccgacggc
     6481 tccctgctgt tccgagtaac catcaacgga gtgaccggct ggcggctgtg cgaacgcatt
     6541 ctggcgtaag aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc
     6601 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggttaa
     6661 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     6721 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     6781 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     6841 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     6901 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     6961 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     7021 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     7081 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     7141 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     7201 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgaa
     7261 acccgctgat cagccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     7321 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     7381 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     7441 tgt
//
