LOCUS       sequence-436565-        6406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of improved GABA sensor (negative change in 
            fluorescence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  AUTHORS   Kolb I, Hasseman JP, Matsumoto A, Arthur BJ, Zhang Y, Tsang A, Reep 
            D, Tsegaye G, Zheng J, Patel R, Looger LL, Marvin JS, Korff WL, 
            Yonehara K, Turner GC
  TITLE     iGABASnFR2: Improved genetically encoded protein sensors of GABA
  JOURNAL   bioRxiv 2025.03.25.644953
REFERENCE   2  (bases 1 to 6406)
  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: "bioRxiv 
            2025.03.25.644953"
COMMENT     Sequence Label: pGP-AAV-GFAP-iGABASnFR2n-WPRE
FEATURES             Location/Qualifiers
     source          1..6406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     regulatory      929..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     935..994
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2041..2062
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2165..2186)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2693..2722
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2726..2872
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2888..3476
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2941..2961)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3359..3370)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3479..3495)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3480..3496)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3525..3646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3536..3555)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3590..3609
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3808..3937
                     /label=AAV2 ITR
     rep_origin      complement(4174..4762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4254..4273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4933..5724,5725..5793))
                     /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"
     promoter        complement(5794..5898)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5924..6379)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6061..6082)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6273..6292
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag atccaacata
      241 tcctggtgtg gagtagggga cgctgctctg acagaggctc gggggcctga gctggctctg
      301 tgagctgggg aggaggcaga cagccaggcc tttgtctgca agcagacctg gcagcattgg
      361 gctggccgcc ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac
      421 acaatgttcg gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc
      481 cttccgagaa gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat
      541 cttgggataa aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg
      601 gtggagaaca aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc
      661 atggacagtg ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac
      721 acaaatgggt gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc
      781 aggctatgcc agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt
      841 cataaagccc tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat
      901 cacctccgct gctcgcgggg atccacccgc caccatggag acagacacac tcctgctatg
      961 ggtactgctg ctctgggttc caggttccac tggtgacaga tctgaaagca tcaatttcgt
     1021 gagctggggc ggtagcaccc aggatgcgca gaagcaggcc tgggccgacc cgttcagcaa
     1081 ggccagcggc attaccgtgg tccaggatgg gcccaccgac tacggcaaac tcaaggccat
     1141 ggtcgaaagc ggcaacgtgc agtgggacgt ggtggatgta gaggccgact tcgccttgcg
     1201 cgccgccgcc gaaggcctgc tcgaacccct ggatttctcg gtgatccagc gcgacaagat
     1261 cgatccgcgg ttcgtgagtg accatggagt gggggccttc ctgttctctc acgtgctcgg
     1321 ctacaacgaa ggcaagctcg gagccagcaa gccccaggac tggaccgcgc tgttcgacac
     1381 caagacctac cccggcaaac gcgccctcta caaatggccg agccctggcg tgctcgaact
     1441 ggccctgctc gccgacggcg taccggccga caagctctac ccgctggacc tggaccgcgc
     1501 cttcaagaaa ctcgacacca tcaagaaaga catcgtctgg tggggcggcg gtgcacagtc
     1561 gcagcagctg ctggcctccg gcgaagtcag catgggccag ttctggaacg gtcgcatcca
     1621 cgccctgcaa gaagacggcg cgccagtggg cgtgagctgg aagcagaacc tggtgatggc
     1681 cgacatcctc gtcgtgccta aaggcacgaa gaacaaagcc gccgcgatga agttcctggc
     1741 cagtgccagc agcgccaaag gccaggacga cttttccgcc ctgaccgcct atgctccagt
     1801 gaacatcgac agtgtgcagc gcctcgacct tgcgcaggtc cggatcaccg ccgacaagca
     1861 gaagaacggc atcatggcga acttcaagat cccgcacaac gtggaggacg gcagcgtgca
     1921 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1981 caaccactac ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca
     2041 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2101 caagggcggt accggaggga gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2161 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg
     2221 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2281 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2341 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2401 ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa
     2461 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2521 cggcaacatc ctggggcaca agctggagta caactggaac gccaacctgg caccgaacct
     2581 gccgaccgcc tacgtcaagg atcagatcac cctcgatttc gcctactggg ccaagaacgg
     2641 tccggccatt gcgacacggt ggaatgaatg gctagtcaaa ctgcaggtcg acgaacaaaa
     2701 actcatctca gaagaggatc tgaatgctgt gggccaggac acgcaggagg tcatcgtggt
     2761 gccacactcc ttgcccttta aggtggtggt gatctcagcc atcctggccc tggtggtgct
     2821 caccatcatc tcccttatca tcctcatcat gctttggcag aagaagccac gttaaaagct
     2881 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2941 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3001 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3061 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3121 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3181 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3241 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3301 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3361 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3421 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3481 ataccgtcga cctcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     3541 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3601 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3661 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     3721 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     3781 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     3841 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3901 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg
     3961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     5941 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6001 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6061 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6121 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6181 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6241 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6301 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6361 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaact
//
