LOCUS       sequence-232418-        5447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arg-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  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 5447)
  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: B5-2
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..166
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     200..219
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        200..218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             258..280
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             290..307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2115..2133)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2129..2176
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2785..3735)
                     /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      3783..4005
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4497..4591)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4622..5416)
                     /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(4733..4752)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5343..5362
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg
       61 cagggcttcc caaccttacc agagggcgcc ccagctggca attccgacgt ctaagaaacc
      121 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
      181 ctcgatgatc ccgcgaaatt aatacgactc actatagggg aattgagagc ggataacaat
      241 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgc atcatcatca
      301 ccatcattta aatattcagg ctcttctctc agaaaaagtc cgtcaggcca tgattgcggc
      361 aggcgcgcct gcggattgcg aaccgcaggt tcgtcagtca gcaaaagttc agttcggcga
      421 ctatcaggct aacggcatga tggcagttgc taaaaaactg ggtatggcac cgcgacaatt
      481 agcagagcag gtgctgactc atctggatct taacggtatc gccagcaaag ttgagatcgc
      541 cggtccaggc tttatcaaca ttttccttga tccggcattc ctggctgaac atgttcagca
      601 ggcgctggcg tccgatcgtc tcggtgttgc tacgccagaa aaacagacca ttgtggttga
      661 ctactctgcg ccaaacgtgg cgaaagagat gcacgtcggt cacctgcgct ctaccattat
      721 tggtgacgca gcagtgcgta ctctggagtt cctcggtcac aaagtgattc gcgcaaacca
      781 cgtcggcgac tggggcactc agttcggtat gctgattgca tggctggaaa agcagcagca
      841 ggaaaacgcc ggtgaaatgg agctggctga ccttgaaggt ttctaccgcg atgcgaaaaa
      901 gcattacgac gaagatgaag agtttgccga gcgtgcacgt aactacgtgg taaaactgca
      961 aagcggtgac gaatatttcc gcgagatgtg gcgcaaactg gtcgacatca ccatgacgca
     1021 gaaccagatc acttacgatc gtctcaacgt gacgctgacc cgtgatgacg tgatgggcga
     1081 aagcctctac aacccaatgc tgccaggaat tgtggcggat ctcaaagcca aaggtctggc
     1141 agtagaaagc gaaggggcga ccgttgtatt ccttgatgag tttaaaaaca aggaaggcga
     1201 accgatgggc gtgatcattc agaagaaaga tggcggctat ctctacacca ccactgatat
     1261 cgcctgtgcg aaatatcgtt atgaaacact gcatgccgat cgcgtgctgt attacatcga
     1321 ctcccgtcag catcaacacc tgatgcaggc atgggcgatc gtccgtaaag caggctatgt
     1381 accggaatcc gtaccgctgg aacaccacat gttcggcatg atgctgggta aagacggcaa
     1441 accgttcaaa acccgcgcgg gtggtacagt gaaactggcc gatctgctgg atgaagccct
     1501 ggaacgtgca cgccgtctgg tggcagaaaa gaacccggat atgccagccg acgagctgga
     1561 aaaactggct aacgcggttg gtattggtgc ggtgaaatat gcggatctct ccaaaaaccg
     1621 caccacggac tacatcttcg actgggacaa catgctggcg tttgagggta ataccgcgcc
     1681 atacatgcag tatgcataca cgcgtgtatt gtccgtgttc cgtaaagcag aaattgacga
     1741 agagcaactg gctgcagctc cggtaatcat ccgtgaagat cgtgaagcgc aactggcagc
     1801 tcgcctgctg cagtttgaag aaaccctcac cgtggttgcc cgtgaaggta cgccgcatgt
     1861 gatgtgtgct tacctgtacg atctggccgg tctgttctct ggcttctacg agcactgccc
     1921 gatcctcagc gcagaaaacg aagaagtgcg taacagccgt ctgaaactgg cacaactgac
     1981 ggcgaagacg ctgaagctgg gtctggatac gctgggtatt gagactgtag agcgtatgta
     2041 ttaaagcaga ggccatatgc tcgaggatcc ggctgctaac aaagcccgaa aggaagctga
     2101 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     2161 cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc
     2221 agtaccggca taaccaagcc tatgcctaca gcatccaggg cctagggtac gggttttgct
     2281 gcccgcaaac gggctgttct ggtgttgcta gtttgttatc agaatcgcag atccggcttc
     2341 agccggtttg ccggctgaaa gcgctatttc ttccagaatt gccatgattt tttccccacg
     2401 ggaggcgtca ctggctcccg tgttgtcggc agctttgatt cgataagcag catcgcctgt
     2461 ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg ttcaatttca
     2521 tgttctagtt gctttgtttt actggtttca cctgttctat taggtgttac atgctgttca
     2581 tctgttacat tgtcgatctg ttcatggtga acagctttga atgcaccaaa aactcgtaaa
     2641 agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga cagttttccc
     2701 tttgatatgt aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg
     2761 atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag
     2821 tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg
     2881 catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc
     2941 gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct gatgtaatgg ttgttggtat
     3001 tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct
     3061 atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa
     3121 tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag
     3181 ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct
     3241 aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata
     3301 aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa
     3361 tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc
     3421 gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt
     3481 cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc
     3541 attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg
     3601 ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag
     3661 cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac
     3721 aactaattca gacatacatc tcaattggtc taggtgattt taatcactat accaattgag
     3781 atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt
     3841 ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag
     3901 acctttgtgt gttttttttg tttatattca agtggttata atttatagaa taaagaaaga
     3961 ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt
     4021 tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta
     4081 aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga atattccttt
     4141 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
     4201 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
     4261 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
     4321 cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg ccctctgatt
     4381 ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc taatgcaccc
     4441 agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtccctagt gcttggattc
     4501 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     4561 gaggtcatta ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg
     4621 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     4681 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     4741 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     4801 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     4861 cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg
     4921 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     4981 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     5041 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     5101 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     5161 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     5221 gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     5281 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     5341 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     5401 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatca
//
