LOCUS       Exported                6716 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.GFAP.iGABASnFR.F102Y.Y137L
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  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 6716)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6716
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        144..174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..198
                     /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)."
     primer_bind     187..209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     206..222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     206..222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   260..389
                     /label=AAV2 ITR
     regulatory      1141..1150
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     1147..1209
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     2253..2274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2377..2398)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2905..2934
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2938..3084
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3100..3688
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3153..3173)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3571..3582)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3691..3707)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3692..3708)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3737..3858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3748..3767)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3802..3821
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4171..4188)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4171..4187)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4330..4785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4417..4436)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4627..4648
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4811..4915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4916..5776
                     /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(5134..5153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5947..6535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6436..6455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagagggagt ggccaactcc atcactaggg gttccttgta gtaatgatta acccagctgg
       61 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      181 attgtgagcg gataacaatt tcacacagga aacagctatg accaggakta cggtgcgggc
      241 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      301 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      361 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      421 atctacgtag ccatgctcta ggaagatcca acatatcctg gtgtggagta ggggacgctg
      481 ctctgacaga ggctcggggg cctgagctgg ctctgtgagc tggggaggag gcagacagcc
      541 aggcctttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      601 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      661 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      721 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      781 ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc
      841 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      901 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      961 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
     1021 gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc
     1081 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggatccc
     1141 gccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc
     1201 actggtgaca gatctgaaag catcaatttc gtgagctggg gcggtagcac ccaggatgcg
     1261 cagaagcagg cctgggccga cccgttcagc aaggccagcg gcattaccgt ggtccaggat
     1321 gggcccaccg actacggcaa actcaaggcc atggtcgaaa gcggcaacgt gcagtgggac
     1381 gtggtggatg tagaggccga cttcgccttg cgcgccgccg ccgaaggcct gctcgaaccc
     1441 ctggatttct cggtgatcca gcgcgacaag atcgacccgc gcttcgtctc cgaccatggc
     1501 gtgggctcgt tcctgtattc tttcgtgctc ggctacaacg aaggcaagct cggagccagc
     1561 aagccccagg actggaccgc gctgttcgac accaagacct accccggcaa acgcgccctc
     1621 ctaaaatggc cgagccctgg cgtgctcgaa ctggccctgc tcgccgacgg cgtaccggcc
     1681 gacaagctct acccgctgga cctggaccgc gccttcaaga aactcgacac catcaagaaa
     1741 gacatcgtct ggtggggcgg cggtgcacag tcgcagcagc tgctggcctc cggcgaagtc
     1801 agcatgggcc agttctggaa cggtcgcatc cacgccctgc aagaagacgg cgcgccagtg
     1861 ggcgtgagct ggaagcagaa cctggtgatg gccgacatcc tcgtcgtgcc taaaggcacg
     1921 aagaacaaag ccgccgcgat gaagttcctg gccagtgcca gcagcgccaa aggccaggac
     1981 gacttttccg ccctgaccgc ctatgctcca gtgaacatcg acagtgtgca gcgcctcgac
     2041 cttgcgcagg tccggatcac cgccgacaag cagaagaacg gcatcatggc gaacttcaag
     2101 atccgccaca acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2161 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgtg
     2221 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2281 gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatgagc
     2341 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2401 aacggccaca agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg
     2461 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2521 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     2581 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatcag cttcaaggac
     2641 gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2701 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2761 tacaactgga acgccaacct ggcaccgaac ctgccgaccg cctacgtcaa ggatcagatc
     2821 accctcgatt tcgcctactg ggccaagaac ggtccggcca ttgcgacacg gtggaatgaa
     2881 tggctagtca aactgcaggt cgacgaacaa aaactcatct cagaagagga tctgaatgct
     2941 gtgggccagg acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg
     3001 gtgatctcag ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc
     3061 atgctttggc agaagaagcc acgttagaag cttatcgata atcaacctct ggattacaaa
     3121 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3181 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3241 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3301 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3361 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3421 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3481 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     3541 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3601 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3661 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg
     3721 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3781 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3841 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3901 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     3961 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     4021 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgactc
     4081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4141 ggcgcgcagc tggcgtaata gcgaagagkc actggccgtc gttttacaac gtcgtgacaa
     4201 aggacacccg gcgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4261 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4321 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4381 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4441 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4501 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4561 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4621 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4681 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4741 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     4801 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4861 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4921 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4981 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5041 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5101 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5161 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5221 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5281 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5341 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5401 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5461 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5521 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5581 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5641 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5701 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5761 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5821 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5881 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6061 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6601 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     6661 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgagcgag cgcgca
//
