LOCUS       sequence-242316-        6566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Val-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6566)
  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 6566)
  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: B1-1
FEATURES             Location/Qualifiers
     source          1..6566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2944..2961
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3035..3053)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3049..3096
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3705..4655)
                     /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      4703..4925
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5417..5511)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5542..6336)
                     /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(5653..5672)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6263..6282
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6515..6533
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taatacgact cactataggg gaattgagag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg catgaaaaga catataaccc acaagatatc
      121 gaacagccgc tttacgagca ctgggaaaag cagggctact ttaagcctaa tggcgatgaa
      181 agccaggaaa gtttctgcat catgatcccg ccgccgaacg tcaccggcag tttgcatatg
      241 ggtcacgcct tccagcaaac catcatggat accatgatcc gctatcagcg catgcagggc
      301 aaaaacaccc tgtggcaggt cggtactgac cacgccggga tcgctaccca gatggtcgtt
      361 gagcgcaaga ttgccgcaga agaaggtaaa acccgtcacg actacggccg cgaagctttc
      421 atcgacaaaa tctgggaatg gaaagcggaa tctggcggca ccattacccg tcagatgcgc
      481 cgtctcggca actccgtcga ctgggagcgt gaacgcttca ccatggacga aggcctgtcc
      541 aatgcggtga aagaagtttt cgttcgtctg tataaagaag acctgattta ccgtggcaaa
      601 cgcctggtaa actgggatcc gaaactgcgc accgctatct ctgacctgga agtcgaaaac
      661 cgcgaatcga aaggttcgat gtggcacatc cgctatccgc tggctgacgg tgcgaaaacc
      721 gcagacggta aagattatct ggtggtcgcg actacccgtc cagaaaccct gctgggcgat
      781 actggcgtag ccgttaaccc ggaagatccg cgttacaaag atctgattgg caaatatgtc
      841 attctgccgc tggttaaccg tcgtattccg atcgttggcg acgaacacgc cgacatggaa
      901 aaaggcaccg gctgcgtgaa aatcactccg gcgcacgact ttaacgacta tgaagtgggt
      961 aaacgtcacg ccctgccgat gatcaacatc ctgacctttg acggcgatat ccgtgaaagc
     1021 gcccaggtgt tcgataccaa aggtaacgaa tctgacgttt attccagcga aatccctgca
     1081 gagttccaga aactggagcg ttttgctgca cgtaaagcag tcgttgccgc agttgacgcg
     1141 cttggcctgc tggaagaaat taaaccgcac gacctgaccg ttccttacgg cgaccgtggc
     1201 ggcgtagtta tcgaaccaat gctgaccgac cagtggtacg tgcgtgccga tgtcctggcg
     1261 aaaccggcgg ttgaagcggt tgagaacggc gacattcagt tcgtaccgaa gcagtacgaa
     1321 aacatgtact tctcctggat gcgcgatatt caggactggt gtatctctcg tcagttgtgg
     1381 tggggtcacc gtatcccggc atggtatgac gaagcgggta acgtttatgt tggccgcaac
     1441 gaagacgaag tgcgtaaaga aaataacctc ggtgctgatg ttgtcctgcg tcaggacgaa
     1501 gacgttctcg atacctggtt ctcttctgcg ctgtggacct tctctaccct tggctggccg
     1561 gaaaataccg acgccctgcg tcagttccac ccaaccagcg tgatggtatc tggtttcgac
     1621 atcattttct tctggattgc ccgcatgatc atgatgacca tgcacttcat caaagatgaa
     1681 aatggcaaac cgcaggtgcc gttccacacc gtttacatga ccggcctgat tcgtgatgac
     1741 gaaggccaga agatgtccaa atccaagggt aacgttatcg acccactgga tatggttgac
     1801 ggtatttcgc tgccagaact gctggaaaaa cgtaccggca atatgatgca gccgcagctg
     1861 gcggacaaaa tccgtaagcg caccgagaag cagttcccga acggtattga gccgcacggt
     1921 actgacgcgc tgcgcttcac cctggcggcg ctggcgtcta ccggtcgtga catcaactgg
     1981 gatatgaagc gtctggaagg ttaccgtaac ttctgtaaca agctgtggaa cgccagccgc
     2041 tttgtgctga tgaacacaga aggtcaggat tgcggcttca acggcggcga aatgacgctg
     2101 tcgctggcgg accgctggat tctggcggag ttcaaccaga ccatcaaagc gtaccgcgaa
     2161 gcgctggaca gcttccgctt cgatatcgcc gcaggcattc tgtatgagtt cacctggaac
     2221 cagttctgtg actggtatct cgagctgacc aagccggtaa tgaacggtgg caccgaagca
     2281 gaactgcgcg gtactcgcca tacgctggtg actgtactgg aaggtctgct gcgcctcgcg
     2341 catccgatca ttccgttcat caccgaaacc atctggcagc gtgtgaaagt actttgcggt
     2401 atcactgccg acaccatcat gctgcagccg ttcccgcagt acgatgcatc tcaggttgat
     2461 gaagccgcac tggccgacac cgaatggctg aaacaggcga tcgttgcggt acgtaacatc
     2521 cgtgcagaaa tgaacatcgc gccgggcaaa ccgctggagc tgctgctgcg tggttgcagc
     2581 gcggatgcag aacgtcgcgt aaatgaaaac cgtggcttcc tgcaaaccct ggcgcgtctg
     2641 gaaagtatca ccgtgctgcc tgccgatgac aaaggtccgg tttccgttac gaagatcatc
     2701 gacggtgcag agctgctgat cccgatggct ggcctcatca acaaagaaga tgagctggcg
     2761 cgtctggcga aagaagtggc gaagattgaa ggtgaaatca gccgtatcga gaacaaactg
     2821 gcgaacgaag gctttgtcgc ccgcgcaccg gaagcggtca tcgcgaaaga gcgtgagaag
     2881 ctggaaggct atgcggaagc gaaagcgaaa ctgattgaac agcaggctgt tatcgccgcg
     2941 ctgcatcatc atcaccatca ttaaagcaga ggccatatgc tcgaggatcc ggctgctaac
     3001 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     3061 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     3121 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
     3181 cctagggtac gggttttgct gcccgcaaac gggctgttct ggtgttgcta gtttgttatc
     3241 agaatcgcag atccggcttc agccggtttg ccggctgaaa gcgctatttc ttccagaatt
     3301 gccatgattt tttccccacg ggaggcgtca ctggctcccg tgttgtcggc agctttgatt
     3361 cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt
     3421 gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat
     3481 taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttga
     3541 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     3601 tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta cttttgtttg
     3661 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     3721 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     3781 ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     3841 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct
     3901 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     3961 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     4021 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     4081 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     4141 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     4201 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     4261 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     4321 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     4381 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     4441 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     4501 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     4561 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     4621 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     4681 taatcactat accaattgag atgggctagt caatgataat tactagtcct tttcctttga
     4741 gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga
     4801 ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca agtggttata
     4861 atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
     4921 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
     4981 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
     5041 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
     5101 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
     5161 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
     5221 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
     5281 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
     5341 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
     5401 tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     5461 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     5521 tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     5581 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     5641 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     5701 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     5761 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
     5821 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     5881 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     5941 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     6001 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     6061 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     6121 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     6181 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     6241 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     6301 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     6361 ttgatcagat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     6421 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtcta
     6481 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6541 tcttcacctc gatgatcccg cgaaat
//
