LOCUS       sequence-433406-        6212 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 6212)
  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 6212)
  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-syn-iGABASnFR2n-WPRE
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        256..703
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      735..744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     741..800
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1847..1868
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1971..1992)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2499..2528
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2532..2678
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2694..3282
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2747..2767)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3165..3176)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3285..3301)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3286..3302)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3331..3452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3342..3361)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3396..3415
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3614..3743
                     /label=AAV2 ITR
     rep_origin      complement(3980..4568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4060..4079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4739..5530,5531..5599))
                     /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     5362..5381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5600..5704)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5730..6185)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5867..5888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6079..6098
                     /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 atctctgcag
      241 agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc
      301 ctacctgacg accgaccccg acccactgga caagcaccca acccccattc cccaaattgc
      361 gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc
      421 agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct
      481 cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca
      541 ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat
      601 aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc
      661 tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta
      721 gcaaggatcc acccgccacc atggagacag acacactcct gctatgggta ctgctgctct
      781 gggttccagg ttccactggt gacagatctg aaagcatcaa tttcgtgagc tggggcggta
      841 gcacccagga tgcgcagaag caggcctggg ccgacccgtt cagcaaggcc agcggcatta
      901 ccgtggtcca ggatgggccc accgactacg gcaaactcaa ggccatggtc gaaagcggca
      961 acgtgcagtg ggacgtggtg gatgtagagg ccgacttcgc cttgcgcgcc gccgccgaag
     1021 gcctgctcga acccctggat ttctcggtga tccagcgcga caagatcgat ccgcggttcg
     1081 tgagtgacca tggagtgggg gccttcctgt tctctcacgt gctcggctac aacgaaggca
     1141 agctcggagc cagcaagccc caggactgga ccgcgctgtt cgacaccaag acctaccccg
     1201 gcaaacgcgc cctctacaaa tggccgagcc ctggcgtgct cgaactggcc ctgctcgccg
     1261 acggcgtacc ggccgacaag ctctacccgc tggacctgga ccgcgccttc aagaaactcg
     1321 acaccatcaa gaaagacatc gtctggtggg gcggcggtgc acagtcgcag cagctgctgg
     1381 cctccggcga agtcagcatg ggccagttct ggaacggtcg catccacgcc ctgcaagaag
     1441 acggcgcgcc agtgggcgtg agctggaagc agaacctggt gatggccgac atcctcgtcg
     1501 tgcctaaagg cacgaagaac aaagccgccg cgatgaagtt cctggccagt gccagcagcg
     1561 ccaaaggcca ggacgacttt tccgccctga ccgcctatgc tccagtgaac atcgacagtg
     1621 tgcagcgcct cgaccttgcg caggtccgga tcaccgccga caagcagaag aacggcatca
     1681 tggcgaactt caagatcccg cacaacgtgg aggacggcag cgtgcagctc gccgaccact
     1741 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1801 gcacccagtc cgtgctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1861 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg
     1921 gagggagcat gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1981 tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca
     2041 ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2101 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2161 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2221 tcagcttcaa ggacgacggc acctacaaga cccgcgccga ggtgaagttc gagggcgaca
     2281 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2341 ggcacaagct ggagtacaac tggaacgcca acctggcacc gaacctgccg accgcctacg
     2401 tcaaggatca gatcaccctc gatttcgcct actgggccaa gaacggtccg gccattgcga
     2461 cacggtggaa tgaatggcta gtcaaactgc aggtcgacga acaaaaactc atctcagaag
     2521 aggatctgaa tgctgtgggc caggacacgc aggaggtcat cgtggtgcca cactccttgc
     2581 cctttaaggt ggtggtgatc tcagccatcc tggccctggt ggtgctcacc atcatctccc
     2641 ttatcatcct catcatgctt tggcagaaga agccacgtta aaagcttatc gataatcaac
     2701 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2761 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2821 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2881 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2941 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3001 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3061 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     3121 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3181 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3241 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     3301 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3361 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3421 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3481 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3541 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3601 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3661 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3721 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3781 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3841 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3901 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3961 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4021 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4081 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4141 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4201 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4261 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4321 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4381 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4441 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4501 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4561 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4621 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4681 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4741 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4801 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4861 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4921 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4981 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5041 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5101 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5161 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5221 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5281 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5341 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5401 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5461 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5521 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5581 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5641 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5701 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     5761 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5821 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5881 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5941 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6001 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6061 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6121 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6181 cgcgtcccat tcgccattca ggctgcgcaa ct
//
