LOCUS       Exported                6905 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Weak affinity glutamate sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CAG-FLEX.SF-iGluSnFR.S72A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Muller 
            JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, 
            Cai E, Hantman AW, Wang SS, DePiero VJ, Borghuis BG, Chapman ER, 
            Dietrich D, DiGregorio DA, Fitzpatrick D, Looger LL
  TITLE     Stability, affinity, and chromatic variants of the glutamate sensor 
            iGluSnFR.
  JOURNAL   Nat Methods. 2018 Nov;15(11):936-939. doi: 
            10.1038/s41592-018-0171-3. Epub 2018 Oct 30.
  PUBMED    30377363
REFERENCE   2  (bases 1 to 6905)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6905
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   65..194
                     /label=AAV2 ITR
     promoter        505..782
                     /label=chicken beta-actin promoter
     primer_bind     1061..1083
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1145..1164
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1199..1232)
                     /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)."
     protein_bind    complement(1295..1328)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1356..1502)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1506..1535)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     2000..2021
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2124..2145)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3111..3173)
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     regulatory      3170..3179
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    3204..3237
                     /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)."
     protein_bind    3308..3341
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3413..4001
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3466..3486)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3884..3895)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4044..4165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4055..4074)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4109..4128
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4327..4456
                     /label=AAV2 ITR
     primer_bind     complement(4522..4539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4693..5281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4773..4792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5452..6312)
                     /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     6075..6094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6313..6417)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6443..6898)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6580..6601)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6792..6811
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
       61 ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
      121 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
      181 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
      241 ctctaggaag agtaccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      481 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      541 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      601 cgatgggggc gggggggggg ggggggcgcg cgccrggsgg ggsggggsgg ggsgrggggs
      661 ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      721 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      781 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      841 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      901 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      961 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg
     1021 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1081 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1141 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccccat
     1201 aacttcgtat aaagtatcct atacgaagtt atatcaaaat aggaagacca atgcttcacc
     1261 atcgacccga attgccaagc atcaccatcg acccataact tcgtataatg tatgctatac
     1321 gaagttatac tagtgctagc ggatccgata tcctaacgtg gcttcttctg ccaaagcatg
     1381 atgaggatga taagggagat gatggtgagc accaccaggg ccaggatggc tgagatcacc
     1441 accaccttaa agggcaagga gtgtggcacc acgatgacct cctgcgtgtc ctggcccaca
     1501 gcattcagat cctcttctga gatgagtttt tgttcgtcga cctgcagttt cagtgccttg
     1561 tcattcggtt ctttgaacag tgctttcatt tcgtctgaca gttcgaaatt catgttcagg
     1621 gggttgttaa agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     1681 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
     1741 gtcttgtagg tgccgtcgtc cttgaagctg atggtgcgct cctggacgta gccttcgggc
     1801 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc
     1861 acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     1921 cagatgaact tcagggtcag cttgccgttg gtggcatcgc cctcgccctc gccgcgcacg
     1981 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     2041 aacagctcct cgcccttgct catgctccct ccggtaccgc ccttgtacag ctcgtccatg
     2101 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     2161 ttggggtctt tgctcagcac ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg
     2221 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     2281 tccacgttgt ggcggatctt gaagttcgcc ttgatgccgt tcttctgctt gtcggcggtg
     2341 atatagacgt tgtggctcac cagaattgga tttttgaacc acttatcaaa ccatttttcc
     2401 gcttcaccgg aggtctgcac ctgagcgatg gtgtcatcca tcagcttttt gaactgcgga
     2461 tcatctttac gcaacataca accgtaggcc tcctgagact gcggcttgcc gacgatttcc
     2521 cagttgtctg gtttcttcgc tttcgcacgt tcaccggcca gcagcacgtc atccatcata
     2581 aaggcaacgg cacgaccgct ttccagggtg cggaaagagt caccgtgatc tttggcgctg
     2641 atgatgcgca tattcatttt ttgctcttca ttcagtttgt tgagcaaaac ttcagaggta
     2701 gtgccggaag tgacgactac ggctttgtct ttcaggttgg caaaatcttt gatatcgcca
     2761 ccctttttgg tcaacaggcg cgtaccgacc acgaaaatag tgtcagagaa agccgcctgt
     2821 ttttggcgtt cgacgttgtt ggtggtagaa ccacattcaa aatcgaaagt gccgttttgc
     2881 agcagtggaa tacggttttg tgcggtaatc ggaatcagtt ttacctgcaa gtccggtttg
     2941 ttgagtttct ttttcactgc ttcaacaatg gcgttggagt aatcctgcga gtaacccacc
     3001 actttttgct gattgtcgta ataagagaaa ggcactgaag attcacggtg accgacgaca
     3061 atcacaccgt ttttggcgat tttgtccagc gtgctgcctg cggcagatct gtcaccagtg
     3121 gaacctggaa cccagagcag cagtacccat agcaggagtg tgtctgtctc catggtggcg
     3181 atatcaccgg tggtaccact agtataactt cgtataggat actttatacg aagttatcat
     3241 tgggattctt cctattttga tccaagcatc accatcgacc ctctagtcca aatctcacca
     3301 tcgacccata acttcgtata gcatacatta tacgaagtta tgtccctcga agaggttcga
     3361 aacactgaaa agggccaatt ctgcaaatga tccactcgag aagcttatcg ataatcaacc
     3421 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3481 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3541 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3601 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3661 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3721 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3781 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3841 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3901 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3961 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacccgg
     4021 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4081 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4141 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4201 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     4261 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     4321 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4381 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4441 gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4501 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5101 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5401 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5461 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5521 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5581 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5641 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5701 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5761 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5821 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5881 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5941 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6001 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6061 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6121 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6181 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6241 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6301 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6361 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6421 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     6481 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     6541 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     6601 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     6661 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     6721 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     6781 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     6841 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     6901 cattc
//
