LOCUS       sequence-130346-        7528 bp    DNA     circular SYN 14-APR-2025
DEFINITION  FLIPQTV3.0 glutamine sensor with R75MY86A mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2012"; volume: "7(6):e38591. doi"; pages: " 
            10.1371/journal.pone.0038591. Epub 2012 Jun 14"
COMMENT     Sequence Label: pcDNA3.1(-) FLIPQTV3.0 8m
FEATURES             Location/Qualifiers
     source          1..7528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(5422..5428,5429..5446,5447..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(join(6532..7323,7324..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 ctacgccaaa 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 aaaaaatggt 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
//
