LOCUS       Exported                6713 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Weak affinity yellow glutamate sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CAG-FLEX.SF-Venus-iGluSnFR.S72A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6713)
  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 6713)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6713
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     413..435
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     497..516
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(551..584)
                     /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(647..680)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(708..854)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(858..887)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1352..1373
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1476..1497)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2463..2525)
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     regulatory      2522..2531
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    2556..2589
                     /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    2660..2693
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2765..3353
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2818..2838)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3236..3247)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3396..3517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3461..3480
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3725..3742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3896..4484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3976..3995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4655..5515)
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5646..6101)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5783..5804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5995..6014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6173..6302
                     /label=AAV2 ITR
ORIGIN
        1 ggggcggggg ggggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
       61 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      121 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      181 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      241 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      301 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
      361 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
      421 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
      481 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
      541 ttggatcccc ataacttcgt ataaagtatc ctatacgaag ttatatcaaa ataggaagac
      601 caatgcttca ccatcgaccc gaattgccaa gcatcaccat cgacccataa cttcgtataa
      661 tgtatgctat acgaagttat actagtgcta gcggatccga tatcctaacg tggcttcttc
      721 tgccaaagca tgatgaggat gataagggag atgatggtga gcaccaccag ggccaggatg
      781 gctgagatca ccaccacctt aaagggcaag gagtgtggca ccacgatgac ctcctgcgtg
      841 tcctggccca cagcattcag atcctcttct gagatgagtt tttgttcgtc gacctgcagt
      901 ttcagtgcct tgtcattcgg ttctttgaac agtgctttca tttcgtctga cagttcgaaa
      961 ttcatgttca gggggttgtt aaagttgtac tccagcttgt gccccaggat gttgccgtcc
     1021 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     1081 acctcggcgc gggtcttgta ggtgccgtcg tccttgaagc tgatggtgcg ctcctggacg
     1141 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     1201 gcgaagcact gcacgccgta gcccagggtg gtcacgaggg tgggccaggg cacgggcagc
     1261 ttgccggtgg tgcagatgag cttcagggtc agcttgccgt tggtggcatc gccctcgccc
     1321 tcgccgcgca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     1381 accaccccgg tgaacagctc ctcgcccttg ctcatgctcc ctccggtacc gcccttgtac
     1441 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
     1501 tcgcgcttct cgttggggtc tttgctcagc acggactgat agctcaggta gtggttgtcg
     1561 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
     1621 acgctgccgt cctccacgtt gtggcggatc ttgaagttcg ccttgatgcc gttcttctgc
     1681 ttgtcggcgg tgatatagac gttgtggctc accagaattg gatttttgaa ccacttatca
     1741 aaccattttt ccgcttcacc ggaggtctgc acctgagcga tggtgtcatc catcagcttt
     1801 ttgaactgcg gatcatcttt acgcaacata caaccgtagg cctcctgaga ctgcggcttg
     1861 ccgacgattt cccagttgtc tggtttcttc gctttcgcac gttcaccggc cagcagcacg
     1921 tcatccatca taaaggcaac ggcacgaccg ctttccaggg tgcggaaaga gtcaccgtga
     1981 tctttggcgc tgatgatgcg catattcatt ttttgctctt cattcagttt gttgagcaaa
     2041 acttcagagg tagtgccgga agtgacgact acggctttgt ctttcaggtt ggcaaaatct
     2101 ttgatatcgc cacccttttt ggtcaacagg cgcgtaccga ccacgaaaat agtgtcagag
     2161 aaagccgcct gtttttggcg ttcgacgttg ttggtggtag aaccacattc aaaatcgaaa
     2221 gtgccgtttt gcagcagtgg aatacggttt tgtgcggtaa tcggaatcag ttttacctgc
     2281 aagtccggtt tgttgagttt ctttttcact gcttcaacaa tggcgttgga gtaatcctgc
     2341 gagtaaccca ccactttttg ctgattgtcg taataagaga aaggcactga agattcacgg
     2401 tgaccgacga caatcacacc gtttttggcg attttgtcca gcgtgctgcc tgcggcagat
     2461 ctgtcaccag tggaacctgg aacccagagc agcagtaccc atagcaggag tgtgtctgtc
     2521 tccatggtgg cgatatcacc ggtggtacca ctagtataac ttcgtatagg atactttata
     2581 cgaagttatc attgggattc ttcctatttt gatccaagca tcaccatcga ccctctagtc
     2641 caaatctcac catcgaccca taacttcgta tagcatacat tatacgaagt tatgtccctc
     2701 gaagaggttc gaaacactga aaagggccaa ttctgcaaat gatccactcg agaagcttat
     2761 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2821 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2881 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2941 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3001 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3061 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3121 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3181 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3241 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3301 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3361 ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3421 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3481 tttatttgta accattataa gctgcaataa acaagttaac aacaagaatt gcattcattt
     3541 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     3601 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     3661 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3721 ctcgctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3781 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3841 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3901 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3961 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4021 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4081 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4141 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4201 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4261 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4321 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4381 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4441 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4501 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4561 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5641 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5701 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5761 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5821 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5881 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5941 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6001 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6061 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     6121 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc
     6181 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     6241 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     6301 cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct ctaggaagag
     6361 taccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6481 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6541 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6601 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     6661 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atg
//
