LOCUS       sequence-307492-        8344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-teminal Flag tagged   aa 1-976 of LARS1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8344)
  AUTHORS   Yoon MS, Son K, Arauz E, Han JM, Kim S, Chen J
  TITLE     Leucyl-tRNA Synthetase Activates Vps34 in Amino Acid-Sensing mTORC1 
            Signaling.
  JOURNAL   Cell Rep. 2016 Aug 9;16(6):1510-1517. doi: 
            10.1016/j.celrep.2016.07.008. Epub 2016 Jul 28.
  PUBMED    27477288
REFERENCE   2  (bases 1 to 8344)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2016.07.008"; journalName: "Cell Rep"; date: 
            "2016-08-9- 9"; volume: "16"; issue: "6"; pages: "1510-1517"
COMMENT     Sequence Label: pcDNA8-Flag-LARS(1-976aa)
FEATURES             Location/Qualifiers
     source          1..8344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      905..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3899..3917)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3900..3917)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3937..3954)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3943..4167
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4213..4641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4300..4319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4510..4531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4650..4670)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4655..4984
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4835..4970
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4897..4916
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5051..5845
                     /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(5105..5124)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5715..5734
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6019..6140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6056..6075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6110..6129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6189..6205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6189..6205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6202..6224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6213..6229
                     /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(6237..6243,6244..6261,6262..6267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6282..6303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6420..6437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6591..7179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6671..6690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7350..8141,8142..8210))
                     /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"
     promoter        complement(8211..8315)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttgggg
      901 ggcggccgcc atggattata aagatgatga tgatggatcg gatcccatgg cggaaagaaa
      961 aggaacagcc aaagtggact ttttgaagaa gattgagaaa gaaatccaac agaaatggga
     1021 tactgagaga gtgtttgagg tcaatgcatc taatttagag aaacagacca gcaagggcaa
     1081 gtattttgta accttcccat atccatatat gaatggacgc cttcatttgg gacacacgtt
     1141 ttctttatcc aaatgtgagt ttgctgtagg gtaccagcga ttgaaaggaa aatgttgtct
     1201 gtttcccttt ggcctgcact gtactggaat gcctattaag gcatgtgctg ataagttgaa
     1261 aagagaaata gagctgtatg gttgcccccc tgattttcca gatgaagaag aggaagagga
     1321 agaaaccagt gttaaaacag aagatataat aattaaggat aaagctaaag gaaaaaagag
     1381 taaagctgct gctaaagctg gatcttctaa ataccagtgg ggcattatga aatcccttgg
     1441 cctgtctgat gaagagatag taaaattttc tgaagcagaa cattggcttg attatttccc
     1501 gccactggct attcaggatt taaaaagaat gggtttgaag gtagactggc gtcgttcctt
     1561 catcaccact gatgttaatc cttactatga ttcatttgtc agatggcaat ttttaacatt
     1621 aagagaaaga aacaaaatta aatttgggaa gcggtataca atttactctc cgaaagatgg
     1681 acagccttgc atggatcatg atagacaaac tggagagggt gttggacctc aggaatatac
     1741 tttactcaaa ttgaaggtgc ttgagccata cccatctaaa ttaagtggcc tgaaaggtaa
     1801 aaatattttc ttggtggctg ctactctcag acctgagacc atgtttgggc agacaaattg
     1861 ttgggttcgt cctgatatga agtacattgg atttgagacg gtgaatggtg atatattcat
     1921 ctgtacccaa aaagcagcca ggaatatgtc ataccagggc tttaccaaag acaatggcgt
     1981 ggtgcctgtt gttaaggaat taatggggga ggaaattctt ggtgcatcac tttctgcacc
     2041 tttaacatca tacaaggtga tctatgttct cccaatgcta actattaagg aggataaagg
     2101 cactggtgtg gttacaagtg ttccttccga ctcccctgat gatattgctg ccctcagaga
     2161 cttgaagaaa aagcaagcct tacgagcaaa atatggaatt agagatgaca tggtcttgcc
     2221 atttgagccg gtgccagtca ttgaaatccc aggttttgga aatctttctg ctgtaaccat
     2281 ttgtgatgag ttgaaaattc agagccagaa tgaccgggaa aaacttgcag aagcaaagga
     2341 gaagatatat ctaaaaggat tttatgaggg tatcatgttg gtggatggat ttaaaggaca
     2401 gaaggttcaa gatgtaaaga agactattca gaaaaagatg attgacgctg gagatgcact
     2461 tatttacatg gaaccagaga aacaagtgat gtccaggtcg tcagatgaat gtgttgtggc
     2521 tctgtgtgac cagtggtact tggattatgg agaagagaat tggaagaaac agacatctca
     2581 gtgcttgaag aacctggaaa cattctgtga ggagaccagg aggaattttg aagccacctt
     2641 aggttggcta caagaacatg cttgctcaag aacttatggt ctaggcactc acctgccttg
     2701 ggatgagcag tggctgattg aatcactttc tgactccact atttacatgg cattttacac
     2761 agttgcacac ctattgcagg ggggtaactt gcatggacag gcagagtctc cgctgggcat
     2821 tagaccgcaa cagatgacca aggaagtttg ggattatgtt ttcttcaagg aggctccatt
     2881 tcctaagact cagattgcaa aggaaaaatt agatcagtta aagcaggagt ttgaattctg
     2941 gtatcctgtt gatcttcgcg tctctggcaa ggatcttgtt ccaaatcatc tttcatatta
     3001 cctttataat catgtggcta tgtggccgga acaaagtgac aaatggccta cagctgtgag
     3061 agcaaatgga catctcctcc tgaactctga gaagatgtca aaatccacag gcaacttcct
     3121 cactttgacc caagctattg acaaattttc agcagatgga atgcgtttgg ctctggctga
     3181 tgctggtgac actgtagaag atgccaactt tgtggaagcc atggcagatg caggtattct
     3241 ccgtctgtac acctgggtag agtgggtgaa agaaatggtt gccaactggg acagcctaag
     3301 aagtggtcct gccagcactt tcaatgatag agtttttgcc agtgaattga atgcaggaat
     3361 tataaaaaca gatcaaaact atgaaaagat gatgtttaaa gaagctttga aaacagggtt
     3421 ttttgagttt caggccgcaa aagataagta ccgtgaattg gctgtggaag ggatgcacag
     3481 agaacttgtg ttccggttta ttgaagttca gacacttctc ctcgctccat tctgtccaca
     3541 tttgtgtgag cacatctgga cactcctggg aaagcctgac tcaattatga atgcttcatg
     3601 gcctgtggca ggtcctgtta atgaagtttt aatacactcc tcacagtatc ttatggaagt
     3661 aacacatgac cttagactac gactcaagaa ctatatgatg ccagctaaag ggaagaagac
     3721 tgacaaacaa cccctgcaga agccctcaca ttgcaccatc tatgtggcaa agaactatcc
     3781 accttggcaa cataccaccc tgtctgttct acgtaaacac tttgaggcca ataacggaaa
     3841 actgcctgac aacaaagtca ttgctagtga actataactc gagtctagag ggccctattc
     3901 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     3961 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4021 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4081 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4141 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4201 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4261 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4321 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4381 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4441 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4501 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4561 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4621 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4681 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4741 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4801 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4861 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4921 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4981 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     5041 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5101 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5161 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5221 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5281 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5341 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5401 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5461 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5521 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5581 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5641 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5701 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5761 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5821 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5881 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5941 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6001 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6061 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6121 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6181 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6241 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6301 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6361 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6421 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6481 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6541 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6601 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6661 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6721 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6781 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6841 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6901 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6961 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7021 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7081 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7141 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7201 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7261 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7321 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7381 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7441 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7501 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7561 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7621 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7681 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7741 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7801 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7861 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7921 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7981 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8041 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8101 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8161 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8221 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8281 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8341 gacg
//
