LOCUS       sequence-292315-        6496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-teminal Flag tagged aa 1-360 of LARS1 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  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 6496)
  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(1-360aa)
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1815..1833)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1816..1833)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1853..1870)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1859..2083
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2129..2557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2216..2235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2426..2447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2566..2586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2571..2900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2751..2886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2813..2832
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2967..3761
                     /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(3021..3040)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3631..3650
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3935..4056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3972..3991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4026..4045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4105..4121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4105..4121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4118..4140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4129..4145
                     /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(4153..4159,4160..4177,4178..4183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4198..4219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4336..4353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4507..5095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4587..4606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5266..6057,6058..6126))
                     /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     5889..5908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6127..6231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6306..6325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggggggcggc cgccatggat tataaagatg atgatgatgg atcggatccc atggcggaaa
      721 gaaaaggaac agccaaagtg gactttttga agaagattga gaaagaaatc caacagaaat
      781 gggatactga gagagtgttt gaggtcaatg catctaattt agagaaacag accagcaagg
      841 gcaagtattt tgtaaccttc ccatatccat atatgaatgg acgccttcat ttgggacaca
      901 cgttttcttt atccaaatgt gagtttgctg tagggtacca gcgattgaaa ggaaaatgtt
      961 gtctgtttcc ctttggcctg cactgtactg gaatgcctat taaggcatgt gctgataagt
     1021 tgaaaagaga aatagagctg tatggttgcc cccctgattt tccagatgaa gaagaggaag
     1081 aggaagaaac cagtgttaaa acagaagata taataattaa ggataaagct aaaggaaaaa
     1141 agagtaaagc tgctgctaaa gctggatctt ctaaatacca gtggggcatt atgaaatccc
     1201 ttggtctgtc tgatgaagag atagtaaaat tttctgaagc agaacattgg cttgattatt
     1261 tcccgccact ggctattcag gatttaaaaa gaatgggttt gaaggtagac tggcgtcgtt
     1321 ccttcatcac cactgatgtt aatccttact atgattcctt tgtcagatgg caatttttaa
     1381 cattaagaga aagaaacaaa attaaatttg ggaagcggta tacaatttac tctccgaaag
     1441 atggacagcc ttgcatggat catgatagac aaactggaga gggtgttgga cctcaggaat
     1501 atactttact caaattgaag gtgcttgagc catacccatc taaattaagt ggcctgaaag
     1561 gtaaaaatat tttcttggtg gctgctactc tcagacctga gaccatgttt gggcagacaa
     1621 attgttgggt tcgtcctgat atgaagtaca ttggatttga gacggtgaat ggtgatatat
     1681 tcatctgtac ccaaaaagca gccaggaata tgtcatacca gggctttacc aaagacaatg
     1741 gcgtggtgcc tgttgttaag gaattaatgg gggaggaaat tcttggtgca taactcgagt
     1801 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     1861 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1921 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1981 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2041 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2101 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2161 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2221 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2281 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2341 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2401 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2461 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2521 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2581 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2641 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2761 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2821 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2881 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2941 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3001 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3061 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3121 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3181 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3241 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3301 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3361 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3421 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3481 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3541 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3601 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3661 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3721 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3781 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3841 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3901 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3961 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4021 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4081 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4141 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4201 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4261 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4321 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4381 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4441 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4921 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5161 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5221 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5281 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5341 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5401 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5461 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5521 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5581 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5641 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5701 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5761 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5821 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5881 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5941 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6001 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6061 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6121 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6181 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6241 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     6301 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     6361 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     6421 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     6481 gccagatata cgcgtt
//
