LOCUS       sequence-307493-        8605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-teminal Flag tagged   aa 1- 1064 of LARS1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8605)
  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 8605)
  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-DELTA-UNE-L
FEATURES             Location/Qualifiers
     source          1..8605
                     /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(4160..4178)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4161..4178)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4198..4215)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4204..4428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4474..4902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4561..4580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4771..4792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4911..4931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4916..5245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5096..5231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5158..5177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5312..6106
                     /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(5366..5385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5976..5995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6280..6401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6317..6336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6371..6390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6450..6466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6450..6466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6463..6485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6474..6490
                     /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(6498..6504,6505..6522,6523..6528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6543..6564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6681..6698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6852..7440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6932..6951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7611..8402,8403..8471))
                     /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(8472..8576)
                     /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 tctgtctgat gaagagatag taaaattttc tgaagcagaa cattggcttg attatttccc
     1501 gccactggct attcaggatt taaaaagaat gggtttgaag gtagactggc gtcgttcctt
     1561 catcaccact gatgttaatc cttactatga ttcctttgtc 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 actaggcagt atgccagaac tgaagaaata
     3901 catgaagaaa gtcatgccat ttgttgccat gattaaggaa aatctggaga agatggggcc
     3961 tcgtattctg gatttgcaat tagaatttga tgaaaaggct gtgcttatgg agaatatagt
     4021 ctatctgact aattcgcttg agctagaaca catagaagtc aagtttgcct ccgaagcaga
     4081 agataaaatc agggaagact gctgtcctgg gaaaccactt aatgttttta gaatataact
     4141 cgagtctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     4201 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4261 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     4321 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     4381 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     4441 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4501 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4561 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4621 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4681 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4741 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4801 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4861 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4921 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4981 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5041 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5101 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5161 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5221 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5281 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5341 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5401 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5461 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5521 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5581 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5641 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5701 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5761 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5821 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5881 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5941 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6001 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6061 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6121 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6181 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6241 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6301 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6361 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6421 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6481 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6541 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6601 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6661 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7261 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8581 tttccccgaa aagtgccacc tgacg
//
