LOCUS       sequence-307495-        7576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-teminal Flag tagged aa 1- 720 of LARS1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7576)
  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 7576)
  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(1-720aa)
FEATURES             Location/Qualifiers
     source          1..7576
                     /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"
     regulatory      903..912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3129..3147)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3130..3147)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3167..3184)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3173..3397
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3443..3871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3530..3549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3740..3761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3880..3900)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3885..4214
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4065..4200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4127..4146
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4281..5075
                     /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(4335..4354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4945..4964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5249..5370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5286..5305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5340..5359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5419..5435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5419..5435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5432..5454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5443..5459
                     /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(5467..5473,5474..5491,5492..5497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5512..5533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5650..5667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5821..6409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5901..5920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6580..7371,7372..7440))
                     /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(7441..7545)
                     /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 gcttgggggg
      901 cggccgccat ggattataaa gatgatgatg atggatcgga tcccatggcg gaaagaaaag
      961 gaacagccaa agtggacttt ttgaagaaga ttgagaaaga aatccaacag aaatgggata
     1021 ctgagagagt gtttgaggtc aatgcatcta atttagagaa acagaccagc aagggcaagt
     1081 attttgtaac cttcccatat ccatatatga atggacgcct tcatttggga cacacgtttt
     1141 ctttatccaa atgtgagttt gctgtagggt accagcgatt gaaaggaaaa tgttgtctgt
     1201 ttccctttgg cctgcactgt actggaatgc ctattaaggc atgtgctgat aagttgaaaa
     1261 gagaaataga gctgtatggt tgcccccctg attttccaga tgaagaagag gaagaggaag
     1321 aaaccagtgt taaaacagaa gatataataa ttaaggataa agctaaagga aaaaagagta
     1381 aagctgctgc taaagctgga tcttctaaat accagtgggg cattatgaaa tcccttggcc
     1441 tgtctgatga agagatagta aaattttctg aagcagaaca ttggcttgat tatttcccgc
     1501 cactggctat tcaggattta aaaagaatgg gtttgaaggt agactggcgt cgttccttca
     1561 tcaccactga tgttaatcct tactatgatt catttgtcag atggcaattt ttaacattaa
     1621 gagaaagaaa caaaattaaa tttgggaagc ggtatacaat ttactctccg aaagatggac
     1681 agccttgcat ggatcatgat agacaaactg gagagggtgt tggacctcag gaatatactt
     1741 tactcaaatt gaaggtgctt gagccatacc catctaaatt aagtggcctg aaaggtaaaa
     1801 atattttctt ggtggctgct actctcagac ctgagaccat gtttgggcag acaaattgtt
     1861 gggttcgtcc tgatatgaag tacattggat ttgagacggt gaatggtgat atattcatct
     1921 gtacccaaaa agcagccagg aatatgtcat accagggctt taccaaagac aatggcgtgg
     1981 tgcctgttgt taaggaatta atgggggagg aaattcttgg tgcatcactt tctgcacctt
     2041 taacatcata caaggtgatc tatgttctcc caatgctaac tattaaggag gataaaggca
     2101 ctggtgtggt tacaagtgtt ccttccgact cccctgatga tattgctgcc ctcagagact
     2161 tgaagaaaaa gcaagcctta cgagcaaaat atggaattag agatgacatg gtcttgccat
     2221 ttgagccggt gccagtcatt gaaatcccag gttttggaaa tctttctgct gtaaccattt
     2281 gtgatgagtt gaaaattcag agccagaatg accgggaaaa acttgcagaa gcaaaggaga
     2341 agatatatct aaaaggattt tatgagggta tcatgttggt ggatggattt aaaggacaga
     2401 aggttcaaga tgtaaagaag actattcaga aaaagatgat tgacgctgga gatgcactta
     2461 tttacatgga accagagaaa caagtgatgt ccaggtcgtc agatgaatgt gttgtggctc
     2521 tgtgtgacca gtggtacttg gattatggag aagagaattg gaagaaacag acatctcagt
     2581 gcttgaagaa cctggaaaca ttctgtgagg agaccaggag gaattttgaa gccaccttag
     2641 gttggctaca agaacatgct tgctcaagaa cttatggtct aggcactcac ctgccttggg
     2701 atgagcagtg gctgattgaa tcactttctg actccactat ttacatggca ttttacacag
     2761 ttgcacacct attgcagggg ggtaacttgc atggacaggc agagtctccg ctgggcatta
     2821 gaccgcaaca gatgaccaag gaagtttggg attatgtttt cttcaaggag gctccatttc
     2881 ctaagactca gattgcaaag gaaaaattag atcagttaaa gcaggagttt gaattctggt
     2941 atcctgttga tcttcgcgtc tctggcaagg atcttgttcc aaatcatctt tcatattacc
     3001 tttataatca tgtggctatg tggccggaac aaagtgacaa atggcctaca gctgtgagag
     3061 caaatggaca tctcctcctg aactctgaga agatgtcaaa atcctaactc gagtctagag
     3121 ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct
     3181 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3241 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3301 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3361 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3421 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3481 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3541 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3601 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3661 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3721 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3781 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3841 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3901 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3961 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4081 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4141 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4201 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4261 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4321 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4381 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4441 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4501 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4561 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4621 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4681 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4741 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4801 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4861 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4921 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4981 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5041 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5101 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5161 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5221 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5341 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5401 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5461 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5521 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5581 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5641 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5701 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7561 agtgccacct gacgtc
//
