LOCUS       Exported                7101 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.hSyn-FLEX.iGABASnFR.F102Y.Y137L
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7101)
  AUTHORS   Marvin JS, Shimoda Y, Magloire V, Leite M, Kawashima T, Jensen TP, 
            Kolb I, Knott EL, Novak O, Podgorski K, Leidenheimer NJ, Rusakov DA,
            Ahrens MB, Kullmann DM, Looger LL
  TITLE     A genetically encoded fluorescent sensor for in vivo imaging of 
            GABA.
  JOURNAL   Nat Methods. 2019 Aug;16(8):763-770. doi: 10.1038/s41592-019-0471-2.
            Epub 2019 Jul 15.
  PUBMED    31308547
REFERENCE   2  (bases 1 to 7101)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7101
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(942..1802)
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1933..2388)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2070..2091)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2282..2301
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2516..2538
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     repeat_region   2566..2695
                     /label=AAV2 ITR
     polyA_signal    2857..2978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2894..2913)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2948..2967
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3056..3644
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3162..3173
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3571..3591
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(3716..3749)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(3820..3853)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     regulatory      3878..3887
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     3884..3946
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     4990..5011
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5114..5135)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5642..5671
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5675..5821
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    5849..5882
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    5945..5978
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     primer_bind     complement(5989..6008)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5991..6123
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   6716..6845
                     /label=AAV2 ITR
     primer_bind     complement(6883..6899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6883..6899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6896..6918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6907..6923
                     /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(6931..6961)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6976..6997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     1981 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2041 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2101 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2161 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2221 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2281 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2341 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     2401 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     2461 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgasggcagg gttttcccag
     2521 tcacgacgtt gtaaaacgac ggccagggcc tcttcgttat tacggctgcg cgctcgctcg
     2581 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     2641 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
     2701 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccttatc
     2761 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca tctccccctg
     2821 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2881 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2941 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg
     3001 cccgggtcga ctctagatgc atgctcgagc ggccgcccgg gtcgacggta tcgatgcggg
     3061 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
     3121 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
     3181 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccaaggaaag
     3241 gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     3301 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
     3361 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
     3421 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
     3481 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     3541 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     3601 ataccagtca atctttcaca aattttgtaa tccagaggtt gattatcgat aagcttctcg
     3661 agtggatcat ttgcagaatt ggcccttttc agtgtttcga acctcttcga gggacataac
     3721 ttcgtataat gtatgctata cgaagttatg ggtcgatggt gagatttgga ctagagggtc
     3781 gatggtgatg cttggatcaa aataggaaga atcccaatga taacttcgta taaagtatcc
     3841 tatacgaagt tatactagtg gtaccaccgg tgatatcgcc accatggaga cagacacact
     3901 cctgctatgg gtactgctgc tctgggttcc aggttccact ggtgacagat ctgaaagcat
     3961 caatttcgtg agctggggcg gtagcaccca ggatgcgcag aagcaggcct gggccgaccc
     4021 gttcagcaag gccagcggca ttaccgtggt ccaggatggg cccaccgact acggcaaact
     4081 caaggccatg gtcgaaagcg gcaacgtgca gtgggacgtg gtggatgtag aggccgactt
     4141 cgccttgcgc gccgccgccg aaggcctgct cgaacccctg gatttctcgg tgatccagcg
     4201 cgacaagatc gacccgcgct tcgtctccga ccatggcgtg ggctcgttcc tgtattcttt
     4261 cgtgctcggc tacaacgaag gcaagctcgg agccagcaag ccccaggact ggaccgcgct
     4321 gttcgacacc aagacctacc ccggcaaacg cgccctccta aaatggccga gccctggcgt
     4381 gctcgaactg gccctgctcg ccgacggcgt accggccgac aagctctacc cgctggacct
     4441 ggaccgcgcc ttcaagaaac tcgacaccat caagaaagac atcgtctggt ggggcggcgg
     4501 tgcacagtcg cagcagctgc tggcctccgg cgaagtcagc atgggccagt tctggaacgg
     4561 tcgcatccac gccctgcaag aagacggcgc gccagtgggc gtgagctgga agcagaacct
     4621 ggtgatggcc gacatcctcg tcgtgcctaa aggcacgaag aacaaagccg ccgcgatgaa
     4681 gttcctggcc agtgccagca gcgccaaagg ccaggacgac ttttccgccc tgaccgccta
     4741 tgctccagtg aacatcgaca gtgtgcagcg cctcgacctt gcgcaggtcc ggatcaccgc
     4801 cgacaagcag aagaacggca tcatggcgaa cttcaagatc cgccacaacg tggaggacgg
     4861 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4921 gctgcccgac aaccactacc tgagcaccca gtccgtgctg agcaaagacc ccaacgagaa
     4981 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     5041 cgagctgtac aagggcggta ccggagggag catgagcaag ggcgaggagc tgttcaccgg
     5101 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg
     5161 cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac
     5221 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     5281 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     5341 aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc
     5401 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     5461 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactggaacg ccaacctggc
     5521 accgaacctg ccgaccgcct acgtcaagga tcagatcacc ctcgatttcg cctactgggc
     5581 caagaacggt ccggccattg cgacacggtg gaatgaatgg ctagtcaaac tgcaggtcga
     5641 cgaacaaaaa ctcatctcag aagaggatct gaatgctgtg ggccaggaca cgcaggaggt
     5701 catcgtggtg ccacactcct tgccctttaa ggtggtggtg atctcagcca tcctggccct
     5761 ggtggtgctc accatcatct cccttatcat cctcatcatg ctttggcaga agaagccacg
     5821 ttaggatatc ggatccgcta gcactagtat aacttcgtat agcatacatt atacgaagtt
     5881 atgtgtcgat ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt cttcctattt
     5941 tgatataact tcgtatagga tactttatac gaagttatgg ggatcatcac ctgtggagag
     6001 aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca agagtcttct
     6061 ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa ccttgatact
     6121 tacctgatcc aacggctgcg acttggggca gggggtccta ggggtggtct ggccaggagc
     6181 cgcgggggcg gactgcgcgg tgcccaggag cacagctgcg ctctcaggca cgacacgact
     6241 cctccgctgc ccaccgcaga ctgaggcagc gctgagtcgc cggcgccgca gcgcagatgg
     6301 tcgcgcccgt gcccccctat ctcgcgcctc gcgtggtgcg gtccggctgg gccggcggcg
     6361 gcgcggacgc gaccaaggtg gccgggaagg ggagtttgcg ggggaccggc gagtgacgtc
     6421 agcgcgcctt cagtgctgag gcggcggtgg cgcgcgccgc caggcggggg cgaaggcact
     6481 gtccgcggtg ctgaagctgg cagtgcgcac gcgcctcgcc gcatcctgtt tcccctcccc
     6541 ctctctgata ggggatgcgc aatttgggga atgggggttg ggtgcttgtc cagtgggtcg
     6601 gggtcggtcg tcaatttaaa tactgaaaag ggcgaattcc agcacactgg agctcggatc
     6661 ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa
     6721 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     6781 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     6841 cgcagctgca ttaatggcgc agccaaccct ggcgtaatca tggtcatagc tgtttcctgt
     6901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7021 gtcccagtcg ggcaaccggg cgcgcagctg cattaatgaa tcggccaacg cgcggggaga
     7081 ggcggtttgc gtattgggcg c
//
