LOCUS Exported 7528 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION FLIPQTV3.0 glutamine sensor with R75MW220A mutations. ACCESSION . VERSION . KEYWORDS pcDNA3.1(-) FLIPQTV3.0 2m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7528) AUTHORS Gruenwald K, Holland JT, Stromberg V, Ahmad A, Watcharakichkorn D, Okumoto S TITLE Visualization of glutamine transporter activities in living cells using genetically encoded glutamine sensors. JOURNAL PLoS One. 2012;7(6):e38591. doi: 10.1371/journal.pone.0038591. Epub 2012 Jun 14. PUBMED 22723868 REFERENCE 2 (bases 1 to 7528) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7528 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1059..1766 /codon_start=1 /product="monomeric teal (cyan) fluorescent protein (Ai et al., 2006)" /label=mTFP1 /note="mammalian codon-optimized" /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA VARNSTDGMDELYK" primer_bind complement(2421..2442) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2682..2701) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 2841..3092 /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK" primer_bind 3029..3050 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3122..3139) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3128..3352 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3398..3826 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3485..3504) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3695..3716 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3835..3855) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3840..4169 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4020..4155 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4082..4101 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4236..5030 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4290..4309) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4900..4919 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5204..5325 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5241..5260) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5295..5314 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5374..5390) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5374..5390) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5387..5409) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5398..5414 /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)." promoter complement(5422..5452) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5467..5488 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5605..5622) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5776..6361) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5856..5875) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6532..7392) /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 7155..7174 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7393..7497) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc 961 accacactgg actagtggat ccactcgaga tggcaggctc gaaattagtt gtcgcgacgg 1021 ataccgcctt cgttccgttc gaatttaaac aggccccaat ggtgagcaag ggcgaggaga 1081 ccacaatggg cgtaatcaag cccgacatga agatcaagct gaagatggag ggcaacgtga 1141 atggccacgc cttcgtgatc gagggcgagg gcgagggcaa gccctacgac ggcaccaaca 1201 ccatcaacct ggaggtgaag gagggagccc ccctgccctt ctcctacgac attctgacca 1261 ccgcgttcgc ctacggcaac agggccttca ccaagtaccc cgacgacatc cccaactact 1321 tcaagcagtc cttccccgag ggctactctt gggagcgcac catgaccttc gaggacaagg 1381 gcatcgtgaa ggtgaagtcc gacatctcca tggaggagga ctccttcatc tacgagatac 1441 acctcaaggg cgagaacttc ccccccaacg gccccgtgat gcagaagaag accaccggct 1501 gggacgcctc caccgagagg atgtacgtgc gcgacggcgt gctgaagggc gacgttaagc 1561 acaagctgct gctggagggc ggcggccacc accgcgttga cttcaagacc atctacaggg 1621 ccaagaaggc ggtgaagctg cccgactatc actttgtgga ccaccgcatc gagatcctga 1681 accacgacaa ggactacaac aaggtgaccg tttacgagag cgccgtggcc cgcaactcca 1741 ccgacggcat ggacgagctg tacaaggtga gcgccaaata tgtgggcttt gacgtcgatc 1801 tgtgggctgc catcgctaaa gagctgaagc tggattacga actgaagccg atggatttca 1861 gtgggatcat tccggcactg caaaccaaaa acgtcgatct ggcgctggcg ggcattacca 1921 tcaccgacga gatgaaaaaa gcgatcgatt tctctgacgg ctactacaaa agcggcctgt 1981 tagtgatggt gaaagctaac aataacgatg tgaaaagcgt gaaagatctc gacgggaaag 2041 tggttgctgt gaagagcggt actggctccg ttgattacgc gaaagcaaac atcaaaacta 2101 aagatctgcg tcagttcccg aacatcgata acgcctatat ggaactgggc accaaccgcg 2161 cagacgccgt tctgcacgat acgccaaaca ttctgtactt catcaaaacc gccggtaacg 2221 gtcagttcaa agcggtaggt gactctctgg aagcgcagca atacggtatt gcgttcccga 2281 aaggtagcga cgagctgcgt gacaaagtca acggcgcgtt gaaaaccctg cgcgagaacg 2341 gaacttacaa cgaaatctac aaaaaagcgt tcggtactga accgaaaggc gctgagctgt 2401 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 2461 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct 2521 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc 2581 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 2641 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 2701 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 2761 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 2821 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc 2881 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca 2941 tcggcgacgg ccccgtgctg ctgcccgaca atcactacct gagctaccag tccgccctga 3001 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg 3061 ggatcactct cggcatggac gagctgtaca agtaaaagct taagtttaaa ccgctgatca 3121 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 3181 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 3241 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 3301 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 3361 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 3421 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 3481 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 3541 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 3601 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 3661 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 3721 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 3781 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 3841 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3901 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 3961 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 4021 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 4081 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 4141 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg 4201 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 4261 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 4321 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 4381 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 4441 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 4501 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 4561 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 4621 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 4681 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 4741 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 4801 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 4861 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 4921 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 4981 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct 5041 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 5101 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 5161 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc 5221 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 5281 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc 5341 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 5401 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 5461 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 5521 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 5581 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 5641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 5701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 5761 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 5821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 5881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 5941 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 6001 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 6061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 6121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 6181 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 6241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 6301 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 6361 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 6421 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 6481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 6541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 6601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 6661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 6721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 6781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 6841 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 6901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7141 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 7201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 7261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 7321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 7381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 7441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 7501 atttccccga aaagtgccac ctgacgtc //