LOCUS       sequence-434364-        7877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of improved GABA sensor (positive change in 
            fluorescence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7877)
  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 7877)
  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-CAG-iGABASnFR2-WPRE-bGH-polyA
FEATURES             Location/Qualifiers
     source          1..7877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1767..1826
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2873..2894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2997..3018)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3525..3554
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3558..3704
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     intron          3773..3905
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3888..3907
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3914..4502
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3967..3987)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4385..4396)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4529..4546)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4535..4646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4742..4762)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4742..4760)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4781..4797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4899..4918)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4964..5019
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5018..5037)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5380..5396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5404..5420
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5428..5434,5435..5452,5453..5458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5473..5494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5553..5748
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5599..5734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5754..5888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5791..5810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5845..5864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5956..5973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6127..6715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6207..6226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6886..7677,7678..7746))
                     /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     7509..7528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7747..7851)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggggg ggcccggggg gggggggggg gggggggggg
      541 gggggggggg gggggggggg gggggggggg ggggccaggg gggggggggg ggggggagag
      601 ttgtgctttt gggggggcgc gggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcggtaccg
     1741 cgggcccggg aaccggtgcc gccaccatgg agacagacac actcctgcta tgggtactgc
     1801 tgctctgggt tccaggttcc actggtgaca gatctgaaag catcaatttc gtgagctggg
     1861 gcggtagcac ccaggatgcg cagaagcagg cctgggccga cccgttcagc aaggccagcg
     1921 gcattaccgt ggtccaggat gggcccaccg actacggcaa actcaaggcc atggtcgaaa
     1981 gcggcaacgt gcagtgggac gtggtggatg tagaggccga cttcgccttg cgcgccgccg
     2041 ccgaaggcct gctcgaaccc ctggatttct cggtgatcca gcgcgacaag atcgatccgc
     2101 ggttcgtgag tgaccatgga gtgggggcct tcctgtactc ttacgtgctc ggctacaacg
     2161 aaggcaagct cggagccagc aagccccagg actggaccgc gctgttcgac accaagacct
     2221 accccggcaa acgcgccctc tacaaatggc cgagccctgg cgtgctcgaa ctggccctgc
     2281 tcgccgacgg cgtaccggcc gacaagctct acccgctgga cctggaccgc gccttcaaga
     2341 aactcgacac catcaagaaa gacatcgtct ggtggggcgg cggtgcacag tcgcagcagc
     2401 tgctggcctc cggcgaagtc agcatgggcc agttctggaa cggtcgcatc cacgccctgc
     2461 aagaagacgg cgcgccagtg ggcgtgagct ggaagcagaa cctggtgatg gccgacatcc
     2521 tcgtcgtgcc taaaggcacg aagaacaaag ccgccgcgat gaagttcctg gccagtgcca
     2581 gcagcgccaa aggccaggac gacttttccg ccctgaccgc ctatgctcca gtgaacatcg
     2641 acagtgtgca gcgcctcgac cttgcgcagg tccggatcac cgccgacaag cagaagaacg
     2701 gcatcatggc gaacttcaag atccgccaca acgtggagga cggcagcgtg cagagcgccg
     2761 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2821 acctgagcac ccagtccgtg ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2881 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
     2941 gtaccggagg gagcatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     3001 tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag ggcgagggcg
     3061 atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     3121 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     3181 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     3241 gcaccatcag cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg
     3301 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     3361 tcctggggca caagctggag tacaactgga acgccaacct ggcaccgaac ctgccgaccg
     3421 cctacgtcaa ggatcagatc accctcgatt tcgcctactg ggccaagaac ggtccggcca
     3481 ttgcgacacg gtggaatgaa tggctagtca aactgcaggt cgacgaacaa aaactcatct
     3541 cagaagagga tctgaatgct gtgggccagg acacgcagga ggtcatcgtg gtgccacact
     3601 ccttgccctt taaggtggtg gtgatctcag ccatcctggc cctggtggtg ctcaccatca
     3661 tctcccttat catcctcatc atgctttggc agaagaagcc acgttaggaa ttcgatatcg
     3721 gtaccgatga atagctcgag gtaccgatga atagctaagg tcgaggccgc aggtaagtat
     3781 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
     3841 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
     3901 cacaggtgtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3961 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4021 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4081 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4141 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4201 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4261 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4321 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4381 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4441 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4501 gcctggaatt cgagctcggt acgatcagcc tcgactgtgc cttctagttg ccagccatct
     4561 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4621 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4681 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagccc agcttttgtt
     4741 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4801 gaaattgtta tccgctaatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     4861 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     4921 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     4981 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     5041 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     5101 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     5161 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     5221 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     5281 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     5341 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5401 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5461 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5521 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     5581 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5641 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5701 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     5761 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5821 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5881 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5941 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6001 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6061 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6121 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6181 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6241 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6301 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6361 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6421 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6481 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6541 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6601 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6661 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6721 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6781 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6841 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6961 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7081 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7321 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7381 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7561 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7741 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7801 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7861 ccgaaaagtg ccacctg
//
