LOCUS       Exported                6828 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Fast indicator for extracellular glutamate imaging.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV iGluf
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Helassa N, Durst CD, Coates C, Kerruth S, Arif U, Schulze C, Wiegert
            JS, Geeves M, Oertner TG, Torok K
  TITLE     Ultrafast glutamate sensors resolve high-frequency release at 
            Schaffer collateral synapses.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 May 7. pii: 1720648115. doi: 
            10.1073/pnas.1720648115.
  PUBMED    29735711
REFERENCE   2  (bases 1 to 6828)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6828
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        46..425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        426..629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     580..600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     674..693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        674..692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     773..835
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     1801..1822
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1928..1949)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2189..2208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2414..2443
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2447..2593
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2615..2632)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2621..2845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2977..3565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3057..3076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3894..3941
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3966..3985
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4173..4967)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4284..4303)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4894..4913
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5066..5085)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5312..5332
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(5415..6275)
                     /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     6038..6057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6276..6380)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6407..34
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6494..6513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6704..6725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgaatttta acaaaatatt aacgcttaca atttacgcgc gcgttgacat tgattattga
       61 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      121 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      301 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      361 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      421 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      541 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      601 tacggtggga ggtctatata agcagaactc tctggctaac tagagaaccc actgcttact
      661 ggcttatcga aattaatacg actcactata gggagaccca agcttggtac cgaactcgga
      721 tccactagta acggccgcca gtgtgctgga attcggcttg gggatatcca ccatggagac
      781 agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg gtgacagatc
      841 tgccgcaggc agcacgctgg acaaaatcgc caaaaacggt gtgattgtcg tcggtcaccg
      901 tgattcttca gtgcctttct cttattacga caatcagcaa aaagtggtgg gttactcgca
      961 ggattactcc aacgccattg ttgaagcagt gaaaaagaaa ctcaacaaac cggacttgca
     1021 ggtaaaactg attccgatta cctcacaaaa ccgtattcca ctgctgcaaa acggcacttt
     1081 cgattttgaa tgtggttcta ccaccaacaa cgtcgaacgc caaaaacagg cggctttctc
     1141 tgacactatt ttcgtggtcg gtacgcgcct gttgaccaaa aagggtggcg atatcaaaga
     1201 ttttgccaac ctgaaagaca aagccgtagt cgtcacttcc ggcactacct ctgaagtttt
     1261 gctcaacaaa ctgaatgaag agcaaaaaat gaatatgcgc atcatcagcg ccaaagatca
     1321 cggtgactct ttccgcaccc tggaaagcgg tcgtgccgtt gcctttatga tggatgacgt
     1381 gctgctggcc ggtgaacgtg cgaaagcgaa gaaaccagac aactgggaaa tcgtcggcaa
     1441 gccgcagtct caggaggcct acggttgtat gttgcgtaaa gatgatccgc agttcaaaaa
     1501 gctgatggat gacaccatcg ctcaggtgca gacctccggt gaagcggaaa aatggtttga
     1561 taagtggttc aaaaatccaa ttctggtgag ccacaacgtc tatatcatgg ccgacaagca
     1621 gaggaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     1681 gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1741 caaccactac ctgagcaccc agtccaaact ttcgaaagac cccaacgaga agcgcgatca
     1801 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1861 caagggcggt accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt
     1921 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1981 gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2041 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2101 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2161 catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2221 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2281 ggacggcaac atcctggggc ataagctgga gtacaacttt aacaaccccc tgaacatgaa
     2341 tttcgaactg tcagacgaaa tgaaagcact gttcaaagaa ccgaatgaca aggcactgaa
     2401 actgcaggtc gacgaacaaa aactcatctc agaagaggat ctgaatgctg tgggccagga
     2461 cacgcaggag gtcatcgtgg tgccacactc cttgcccttt aaggtggtgg tgatctcagc
     2521 catcctggcc ctggtggtgc tcaccatcat ctcccttatc atcctcatca tgctttggca
     2581 gaagaagcca cgttaggcgg ccgctcgaga tcagcctcga ctgtgccttc tagttgccag
     2641 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2701 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2761 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcgt
     2821 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagtgg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3121 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3361 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3721 acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc
     3781 cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg
     3841 tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa
     3901 cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
     3961 cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag
     4021 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     4081 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
     4141 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     4201 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4261 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4321 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4381 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcttgctc
     4441 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4501 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     4561 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4621 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4681 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4741 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
     4801 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4861 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     4921 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     4981 catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     5041 cttctggaat agctcagagg ccgggcctcg gcctctgcat aaataaaaaa aattagtcag
     5101 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     5161 ggcgggacta tggttgctga ctaattgaga tgcttgcttt gcatacttct gcctgctggg
     5221 gagcctgggg actttccaca cctggttgct gactaattga gatgcttgct ttgcatactt
     5281 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     5341 ttctttccgc ctcaggactc ttcctttttc aataaatcaa tctaaagtat atatgagtaa
     5401 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5461 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5521 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5581 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5641 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5701 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5761 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5821 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5881 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5941 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6001 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6061 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6121 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6181 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6241 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6301 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6361 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     6421 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6481 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6541 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6601 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6661 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6721 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6781 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaac
//
