LOCUS       sequence-433402-        4480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial 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 4480)
  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 4480)
  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-RSET-iGABASnFR2n-6xHis
FEATURES             Location/Qualifiers
     source          1..4480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     1143..1164
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1267..1288)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             1789..1806
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1881..1899)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1895..1942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2039..2494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2126..2145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2336..2357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2520..2624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2625..2693,2694..3485)
                     /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(2843..2862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3656..4244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4145..4164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4398..4415
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgggagaatc gattaatttc
      121 gtgagctggg gcggtagcac ccaggatgcg cagaagcagg cctgggccga cccgttcagc
      181 aaggccagcg gcattaccgt ggtccaggat gggcccaccg actacggcaa actcaaggcc
      241 atggtcgaaa gcggcaacgt gcagtgggac gtggtggatg tagaggccga cttcgccttg
      301 cgcgccgccg ccgaaggcct gctcgaaccc ctggatttct cggtgatcca gcgcgacaag
      361 atcgatccgc ggttcgtgag tgaccatgga gtgggggcct tcctgttctc tcacgtgctc
      421 ggctacaacg aaggcaagct cggagccagc aagccccagg actggaccgc gctgttcgac
      481 accaagacct accccggcaa acgcgccctc tacaaatggc cgagccctgg cgtgctcgaa
      541 ctggccctgc tcgccgacgg cgtaccggcc gacaagctct acccgctgga cctggaccgc
      601 gccttcaaga aactcgacac catcaagaaa gacatcgtct ggtggggcgg cggtgcacag
      661 tcgcagcagc tgctggcctc cggcgaagtc agcatgggcc agttctggaa cggtcgcatc
      721 cacgccctgc aagaagacgg cgcgccagtg ggcgtgagct ggaagcagaa cctggtgatg
      781 gccgacatcc tcgtcgtgcc taaaggcacg aagaacaaag ccgccgcgat gaagttcctg
      841 gccagtgcca gcagcgccaa aggccaggac gacttttccg ccctgaccgc ctatgctcca
      901 gtgaacatcg acagtgtgca gcgcctcgac cttgcgcagg tccggatcac cgccgacaag
      961 cagaagaacg gcatcatggc gaacttcaag atcccgcaca acgtggagga cggcagcgtg
     1021 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1081 gacaaccact acctgagcac ccagtccgtg ctgagcaaag accccaacga gaagcgcgat
     1141 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1201 tacaagggcg gtaccggagg gagcatgagc aagggcgagg agctgttcac cggggtggtg
     1261 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag
     1321 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
     1381 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1441 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1501 gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg cgccgaggtg
     1561 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1621 gacggcaaca tcctggggca caagctggag tacaactgga acgccaacct ggcaccgaac
     1681 ctgccgaccg cctacgtcaa ggatcagatc accctcgatt tcgcctactg ggccaagaac
     1741 ggtccggcca tcgcgacacg gtggaatgaa tggctagtca aaggtagcca tcatcaccat
     1801 caccactagg gacgcgtaag atatcaagct tgatccggct gctaacaaag cccgaaagga
     1861 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1921 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatctg gcgtaatagc
     1981 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     2041 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2101 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2161 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2221 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2281 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2341 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2401 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2461 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     2521 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2581 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2641 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2701 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2761 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2821 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2881 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2941 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3001 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3061 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3121 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3181 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3241 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3301 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3361 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3421 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3481 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3541 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3601 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3661 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3721 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3781 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3841 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3901 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3961 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4021 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4081 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4141 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4201 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4261 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4321 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4381 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4441 aaccgcctct ccccgcgcgt tggccgattc attaatgcag
//
