LOCUS       Exported                6786 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Weak affinity yellow glutamate sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.hSynap-FLEX.SF-Venus-iGluSnFR.S72A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6786)
  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 6786)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6786
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(1126..1159)
                     /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(1222..1255)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1283..1429)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1433..1462)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1927..1948
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2051..2072)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3038..3100)
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     regulatory      3097..3106
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    3131..3164
                     /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    3235..3268
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3340..3928
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3393..3413)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3811..3822)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4006..4127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4017..4036)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4071..4090
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4481..5069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4561..4580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5240..6100)
                     /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     5863..5882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6101..6205)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6231..6686)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6368..6389)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6580..6599
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
       61 gcgncagrgw ggsmgwstgg aamkcsrkca stgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacmmwtt casrcwgsrm ammkstwggr amgggcgawc ggtgcgggcc
      241 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatccga gctccagtgt gctggaattc gcccttttca
      481 gtatttaaat tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa
      541 attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca
      601 ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc
      661 cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc
      721 ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc
      781 gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct
      841 cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagct gtgctcctgg
      901 gcaccgcgca gtccgccccc gcggctcctg gccagaccac ccctaggacc ccctgcccca
      961 agtcgcagcc gttggatcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgatga tccccataac ttcgtataaa
     1141 gtatcctata cgaagttata tcaaaatagg aagaccaatg cttcaccatc gacccgaatt
     1201 gccaagcatc accatcgaca cataacttcg tataatgtat gctatacgaa gttatactag
     1261 tgctagcgga tccgatatcc taacgtggct tcttctgcca aagcatgatg aggatgataa
     1321 gggagatgat ggtgagcacc accagggcca ggatggctga gatcaccacc accttaaagg
     1381 gcaaggagtg tggcaccacg atgacctcct gcgtgtcctg gcccacagca ttcagatcct
     1441 cttctgagat gagtttttgt tcgtcgacct gcagtttcag tgccttgtca ttcggttctt
     1501 tgaacagtgc tttcatttcg tctgacagtt cgaaattcat gttcaggggg ttgttaaagt
     1561 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     1621 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtaggtgc
     1681 cgtcgtcctt gaagctgatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     1741 agaagtcgtg ctgcttcatg tggtcggggt agcgggcgaa gcactgcacg ccgtagccca
     1801 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgagcttca
     1861 gggtcagctt gccgttggtg gcatcgccct cgccctcgcc gcgcacgctg aacttgtggc
     1921 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     1981 ccttgctcat gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc
     2041 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2101 tcagcacgga ctgatagctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
     2161 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcc acgttgtggc
     2221 ggatcttgaa gttcgccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
     2281 ggctcaccag aattggattt ttgaaccact tatcaaacca tttttccgct tcaccggagg
     2341 tctgcacctg agcgatggtg tcatccatca gctttttgaa ctgcggatca tctttacgca
     2401 acatacaacc gtaggcctcc tgagactgcg gcttgccgac gatttcccag ttgtctggtt
     2461 tcttcgcttt cgcacgttca ccggccagca gcacgtcatc catcataaag gcaacggcac
     2521 gaccgctttc cagggtgcgg aaagagtcac cgtgatcttt ggcgctgatg atgcgcatat
     2581 tcattttttg ctcttcattc agtttgttga gcaaaacttc agaggtagtg ccggaagtga
     2641 cgactacggc tttgtctttc aggttggcaa aatctttgat atcgccaccc tttttggtca
     2701 acaggcgcgt accgaccacg aaaatagtgt cagagaaagc cgcctgtttt tggcgttcga
     2761 cgttgttggt ggtagaacca cattcaaaat cgaaagtgcc gttttgcagc agtggaatac
     2821 ggttttgtgc ggtaatcgga atcagtttta cctgcaagtc cggtttgttg agtttctttt
     2881 tcactgcttc aacaatggcg ttggagtaat cctgcgagta acccaccact ttttgctgat
     2941 tgtcgtaata agagaaaggc actgaagatt cacggtgacc gacgacaatc acaccgtttt
     3001 tggcgatttt gtccagcgtg ctgcctgcgg cagatctgtc accagtggaa cctggaaccc
     3061 agagcagcag tacccatagc aggagtgtgt ctgtctccat ggtggcgata tcaccggtgg
     3121 taccactagt ataacttcgt ataggatact ttatacgaag ttatcattgg gattcttcct
     3181 attttgatcc aagcatcacc atcgaccctc tagtccaaat ctcaccatcg acccataact
     3241 tcgtatagca tacattatac gaagttatgt ccctcgaaga ggttcgaaac actgaaaagg
     3301 gccaattctg caaatgatcc actcgagaag cttatcgata atcaacctct ggattacaaa
     3361 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3421 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3481 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3541 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3601 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3661 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3721 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     3781 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3841 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3901 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg gccgctcgag
     3961 catgcatcta gagtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg
     4021 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4081 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4141 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     4201 acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag
     4261 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     4321 tctctgcgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     6241 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     6301 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     6361 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     6421 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     6481 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     6541 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     6601 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6661 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     6721 agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct cgctcactga
     6781 ggccgc
//
