LOCUS       sequence-213286-        5228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lys-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5228)
  AUTHORS   Villarreal F, Contreras-Llano LE, Chavez M, Ding Y, Fan J, Pan T, 
            Tan C
  TITLE     Synthetic microbial consortia enable rapid assembly of pure 
            translation machinery.
  JOURNAL   Nat Chem Biol. 2018 Jan;14(1):29-35. doi: 10.1038/nchembio.2514. 
            Epub 2017 Nov 13.
  PUBMED    29131146
REFERENCE   2  (bases 1 to 5228)
  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: 
            "10.1038/nchembio.2514"; journalName: "Nat Chem Biol"; date: 
            "2018-01"; volume: "14"; issue: "1"; pages: "29-35"
COMMENT     Sequence Label: B2-3
FEATURES             Location/Qualifiers
     source          1..5228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     123..141
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     175..194
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        175..193
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             233..255
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1780..1797
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1871..1889)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1885..1932
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2541..3491)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3539..3761
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4253..4347)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4378..5172)
                     /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(4489..4508)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5099..5118
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg
       61 gcgccccagc tggcaattcc gacgtctaag aaaccattat tatcatgaca ttaacctata
      121 aaaataggcg tatcacgagg ccctttcgtc ttcacctcga tgatcccgcg aaattaatac
      181 gactcactat aggggaattg agagcggata acaattcccc tctagaaata attttgttta
      241 actttaagaa ggagatatac catgcattct gaacaacacg cacagggcgc tgacgcggta
      301 gtcgatctta acaatgaact gaaaacgcgt cgtgagaagc tggcgaacct gcgcgagcag
      361 gggattgcct tcccgaacga tttccgtcgc gatcatacct ctgaccaatt gcacgcagaa
      421 ttcgacggca aagagaacga agaactggaa gcgctgaaca tcgaagtcgc cgttgctggc
      481 cgcatgatga cccgtcgtat tatgggtaaa gcgtctttcg ttaccctgca ggacgttggc
      541 ggtcgcattc agctgtacgt tgcccgtgac gatctcccgg aaggcgttta taacgagcag
      601 ttcaaaaaat gggacctcgg cgacatcctc ggcgcgaaag gtaagctgtt caaaaccaaa
      661 accggcgaac tgtctatcca ctgcaccgag ttgcgtctgc tgaccaaagc actgcgtccg
      721 ctgccggata aattccacgg cttgcaggat caggaagcgc gctatcgtca gcgttatctc
      781 gatctcatct ccaacgatga atcccgcaac acctttaaag tgcgctcgca gatcctctct
      841 ggtattcgcc agttcatggt gaaccgcggc tttatggaag ttgaaacgcc gatgatgcag
      901 gtgatccctg gcggtgccgc tgcgcgtccg tttatcaccc accataacgc gctggatctc
      961 gacatgtacc tgcgtatcgc gccggaactg tacctcaagc gtctggtggt tggtggcttc
     1021 gagcgtgtat tcgaaatcaa ccgtaacttc cgtaacgaag gtatttccgt acgtcataac
     1081 ccagagttca ccatgatgga actctacatg gcttacgcag attacaaaga tctgatcgag
     1141 ctgaccgaat cgctgttccg tactctggca caggatattc tcggtaagac ggaagtgacc
     1201 tacggcgacg tgacgctgga cttcggtaaa ccgttcgaaa aactgaccat gcgtgaagcg
     1261 atcaagaaat atcgcccgga aaccgacatg gcggatctgg acaacttcga ctctgcgaaa
     1321 gcaattgctg aatctatcgg catccacgtt gagaagagct ggggtctggg ccgtatcgtt
     1381 accgagatct tcgaagaagt ggcagaagca catctgattc agccgacctt cattactgaa
     1441 tatccggcag aagtttctcc gctggcgcgt cgtaacgacg ttaacccgga aatcacagac
     1501 cgctttgagt tcttcattgg tggtcgtgaa atcggtaacg gctttagcga gctgaatgac
     1561 gcggaagatc aggcgcaacg cttcctggat caggttgccg cgaaagacgc aggtgacgac
     1621 gaagcgatgt tctacgatga agattacgtc accgcactgg aacatggctt accgccgaca
     1681 gcaggtctgg gaattggtat cgaccgtatg gtaatgctgt tcaccaacag ccataccatc
     1741 cgcgacgtta ttctgttccc ggcgatgcgt ccggtaaaac atcatcatca ccatcattaa
     1801 agcagcggcc atatgctcga ggatccggct gctaacaaag cccgaaagga agctgagttg
     1861 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1921 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     1981 ccggcataac caagcctatg cctacagcat ccagggccta gggtacgggt tttgctgccc
     2041 gcaaacgggc tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc
     2101 ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag
     2161 gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca
     2221 ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt
     2281 ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg
     2341 ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct
     2401 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg
     2461 atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag
     2521 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg
     2581 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg
     2641 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt
     2701 agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg
     2761 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct
     2821 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc
     2881 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt
     2941 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc
     3001 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc
     3061 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt
     3121 tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt
     3181 gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg
     3241 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt
     3301 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct
     3361 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg
     3421 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact
     3481 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg
     3541 gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc
     3601 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct
     3661 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa
     3721 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg
     3781 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac
     3841 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc
     3901 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     3961 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     4021 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     4081 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     4141 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     4201 aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac
     4261 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     4321 tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga gtcccgctca
     4381 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     4441 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     4501 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     4561 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     4621 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     4681 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     4741 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     4801 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     4861 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     4921 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     4981 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     5041 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     5101 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     5161 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg
     5221 ccatcaga
//
