LOCUS       Exported                6840 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Weak affinity yellow glutamate sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CAG.SF-Venus-iGluSnFR.S72A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6840)
  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 6840)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6840
                     /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
     promoter        702..979
                     /label=chicken beta-actin promoter
     primer_bind     1259..1281
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1343..1362
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1396..1405
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     1402..1464
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     2430..2451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2554..2575)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3040..3069
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3073..3219
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3235..3823
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3288..3308)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3706..3717)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3866..3987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3877..3896)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3931..3950
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4149..4278
                     /label=AAV2 ITR
     primer_bind     complement(4297..4314)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4297..4313)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4306..4328)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4455..4910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4542..4561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4752..4773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4936..5040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5041..5901
                     /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(5259..5278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6072..6660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6561..6580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6814..6831
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
       61 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      181 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      241 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      301 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      361 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      421 atctacgtag ccatgctcta ggaagatcgt accattgacg tcaataatga cgtatgttcc
      481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggactatt tacggtaaac
      541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
      721 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
      781 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc
      841 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      901 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      961 gcgaagcgcg cggcgggcgg gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg
     1021 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     1081 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
     1141 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
     1201 gcggctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1261 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     1321 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     1381 aaagaattgg atcccgccac catggagaca gacacactcc tgctatgggt actgctgctc
     1441 tgggttccag gttccactgg tgacagatct gccgcaggca gcacgctgga caaaatcgcc
     1501 aaaaacggtg tgattgtcgt cggtcaccgt gaatcttcag tgcctttctc ttattacgac
     1561 aatcagcaaa aagtggtggg ttactcgcag gattactcca acgccattgt tgaagcagtg
     1621 aaaaagaaac tcaacaaacc ggacttgcag gtaaaactga ttccgattac cgcacaaaac
     1681 cgtattccac tgctgcaaaa cggcactttc gattttgaat gtggttctac caccaacaac
     1741 gtcgaacgcc aaaaacaggc ggctttctct gacactattt tcgtggtcgg tacgcgcctg
     1801 ttgaccaaaa agggtggcga tatcaaagat tttgccaacc tgaaagacaa agccgtagtc
     1861 gtcacttccg gcactacctc tgaagttttg ctcaacaaac tgaatgaaga gcaaaaaatg
     1921 aatatgcgca tcatcagcgc caaagatcac ggtgactctt tccgcaccct ggaaagcggt
     1981 cgtgccgttg cctttatgat ggatgacgtg ctgctggccg gtgaacgtgc gaaagcgaag
     2041 aaaccagaca actgggaaat cgtcggcaag ccgcagtctc aggaggccta cggttgtatg
     2101 ttgcgtaaag atgatccgca gttcaaaaag ctgatggatg acaccatcgc tcaggtgcag
     2161 acctccggtg aagcggaaaa atggtttgat aagtggttca aaaatccaat tctggtgagc
     2221 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc
     2281 cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2341 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctatca gtccgtgctg
     2401 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2461 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catgagcaag
     2521 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2581 ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc
     2641 ctgaagctca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2701 ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2761 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac
     2821 ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2881 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2941 aactttaaca accccctgaa catgaatttc gaactgtcag acgaaatgaa agcactgttc
     3001 aaagaaccga atgacaaggc actgaaactg caggtcgacg aacaaaaact catctcagaa
     3061 gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg
     3121 ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc
     3181 cttatcatcc tcatcatgct ttggcagaag aagccacgtt agaagcttat cgataatcaa
     3241 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3301 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3361 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3421 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3481 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3541 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3601 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3661 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3721 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3781 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc
     3841 gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     3901 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3961 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4021 gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     4081 atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     4141 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4201 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4261 gagcgagcga gcgcgcagcc ttaattaacc taattcactg gccgtcgttt tacaacgtcg
     4321 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     4381 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4441 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4501 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4561 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4621 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4681 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4741 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4801 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4861 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4921 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4981 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5041 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5101 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5161 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5221 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5281 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5341 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5401 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5461 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5521 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5581 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5641 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5701 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5761 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5821 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5881 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5941 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6001 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6061 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6181 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
//
