LOCUS       Exported                6324 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Tight affinity glutamate sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.hSynapsin.SF-iGluSnFR.A184S
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6324)
  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 6324)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6324
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(244..832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(324..343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1003..1863)
                     /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     1626..1645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1864..1968)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1994..2449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2131..2152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2343..2362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2521..2650
                     /label=AAV2 ITR
     promoter        2726..3173
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3202..3211
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     3208..3270
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     4236..4257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4360..4381)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4846..4875
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4879..5025
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    5041..5629
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5094..5114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5512..5523)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5632..5648)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5633..5649)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5678..5799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5689..5708)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5743..5762
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5961..6090
                     /label=AAV2 ITR
ORIGIN
        1 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
       61 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1981 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2041 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2101 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2161 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2221 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2281 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2341 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2401 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     2461 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     2521 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
     2581 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     2641 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
     2701 aggaagatcc agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg
     2761 gtgggggtgc ctacctgacg accggccccg acccactgga caagcaccca acccccattc
     2821 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt
     2881 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc
     2941 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc
     3001 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga
     3061 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc
     3121 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtccaatt
     3181 caagctgcta gcaaggatcc cgccaccatg gagacagaca cactcctgct atgggtactg
     3241 ctgctctggg ttccaggttc cactggtgac agatctgccg caggcagcac gctggacaaa
     3301 atcgccaaaa acggtgtgat tgtcgtcggt caccgtgaat cttcagtgcc tttctcttat
     3361 tacgacaatc agcaaaaagt ggtgggttac tcgcaggatt actccaacgc cattgttgaa
     3421 gcagtgaaaa agaaactcaa caaaccggac ttgcaggtaa aactgattcc gattacctca
     3481 caaaaccgta ttccactgct gcaaaacggc actttcgatt ttgaatgtgg ttctaccacc
     3541 aacaacgtcg aacgccaaaa acaggcggct ttctctgaca ctattttcgt ggtcggtacg
     3601 cgcctgttga ccaaaaaggg tggcgatatc aaagattttg ccaacctgaa agacaaagcc
     3661 gtagtcgtca cttccggcac tacctctgaa gttttgctca acaaactgaa tgaagagcaa
     3721 aaaatgaata tgcgcatcat cagcgccaaa gatcacggtg actctttccg caccctggaa
     3781 agcggtcgtg ccgttgcctt tatgatggat gacagcctgc tggccggtga acgtgcgaaa
     3841 gcgaagaaac cagacaactg ggaaatcgtc ggcaagccgc agtctcagga ggcctacggt
     3901 tgtatgttgc gtaaagatga tccgcagttc aaaaagctga tggatgacac catcgctcag
     3961 gtgcagacct ccggtgaagc ggaaaaatgg tttgataagt ggttcaaaaa tccaattctg
     4021 gtgagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggcgaacttc
     4081 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4141 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4201 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4261 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg
     4321 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     4381 gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag
     4441 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4501 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     4561 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag
     4621 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     4681 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     4741 gagtacaact ttaacaaccc cctgaacatg aatttcgaac tgtcagacga aatgaaagca
     4801 ctgttcaaag aaccgaatga caaggcactg aaactgcagg tcgacgaaca aaaactcatc
     4861 tcagaagagg atctgaatgc tgtgggccag gacacgcagg aggtcatcgt ggtgccacac
     4921 tccttgccct ttaaggtggt ggtgatctca gccatcctgg ccctggtggt gctcaccatc
     4981 atctccctta tcatcctcat catgctttgg cagaagaagc cacgttagaa gcttatcgat
     5041 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5101 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5161 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5221 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5281 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5341 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     5401 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     5461 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5521 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5581 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     5641 cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     5701 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5761 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5821 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     5881 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     5941 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     6001 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     6061 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     6121 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6181 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6241 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6301 cgtaaaaagg ccgcgttgct ggcg
//
