LOCUS       sequence-213277-        4712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trp-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4712)
  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 4712)
  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: B5-4
FEATURES             Location/Qualifiers
     source          1..4712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)"
     primer_bind     complement(1355..1373)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1369..1416
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2025..2975)
                     /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      3023..3245
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3737..3831)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3862..4656)
                     /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(3973..3992)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4583..4602
                     /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 gatcacgagg ccctttcgtc ttcacctcga tgatcccgcg aaattaatac
      181 gactcactat aggggaattg agagcggata acaattcccc tctagaaata attttgttta
      241 actttaagaa ggagatatac catgcatact aagcccatcg tttttagtgg cgcacagccc
      301 tcaggtgaat tgaccattgg taactacatg ggtgcgctgc gtcagtgggt aaacatgcag
      361 gatgactacc attgcattta ctgtatcgtt gaccaacacg cgatcaccgt gcgccaggat
      421 gcacagaagc tgcgtaaagc gacgctggat acgctggcct tgtatctggc ttgtggtatc
      481 gatcctgaga aaagcaccat ttttgttcag tcccacgtgc cggaacatgc acagttaggc
      541 tgggcactga actgctatac ctacttcggc gaactgagtc gcatgacgca gtttaaagat
      601 aaatctgcgc gttatgccga gaacatcaac gctggtctgt ttgactatcc ggtgctgatg
      661 gcagcggaca tcctgctgta tcaaactaat ctggtaccgg tgggtgaaga ccagaaacag
      721 cacctcgaac tgagccgcga tattgcccag cgtttcaacg cgctgtatgg cgagatcttt
      781 aaggtgccgg agccgtttat tccgaaatct ggcgcgcgcg taatgtcgct gctggagccg
      841 accaagaaga tgtccaagtc tgacgataat cgcaataacg ttatcggcct gctggaagat
      901 ccgaaatcgg tagtgaagaa aatcaaacgt gcggtcactg actccgacga gccgccggta
      961 gttcgctacg atgtgcagaa caaagcgggc gtttccaacc tgttggatat cctttcagcg
     1021 gtaacgggcc agagcatccc agaactggaa aaacagttcg aaggcaagat gtatggtcat
     1081 ctgaaaggtg aagtggctga tgccgtttcc ggtatgctga ctgaattgca ggaacgctat
     1141 caccgtttcc gcaacgatga agccttcctg caacaggtga tgaaagatgg cgcggaaaaa
     1201 gccagcgcgc acgcttcccg tacgctaaaa gcggtgtacg aagcgattgg ttttgtggcg
     1261 aagccgcatc atcaccatca ttaaagcaga ggccatatgc tcgaggatcc ggctgctaac
     1321 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1381 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1441 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
     1501 cctagggtac gggttttgct gcccgcaaac gggctgttct ggtgttgcta gtttgttatc
     1561 agaatcgcag atccggcttc agccggtttg ccggctgaaa gcgctatttc ttccagaatt
     1621 gccatgattt tttccccacg ggaggcgtca ctggctcccg tgttgtcggc agctttgatt
     1681 cgataagcag catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt
     1741 gtctcaggtg ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat
     1801 taggtgttac atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttga
     1861 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     1921 tgcatatgga cagttttccc tttgatatgt aacggtgaac agttgttcta cttttgtttg
     1981 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     2041 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     2101 ttgagatcat acttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     2161 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct
     2221 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     2281 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     2341 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     2401 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     2461 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     2521 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     2581 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     2641 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     2701 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     2761 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     2821 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     2881 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     2941 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     3001 taatcactat accaattgag atgggctagt caatgataat tactagtcct tttcctttga
     3061 gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga
     3121 ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca agtggttata
     3181 atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
     3241 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
     3301 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
     3361 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
     3421 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
     3481 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
     3541 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
     3601 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
     3661 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
     3721 tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     3781 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     3841 tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     3901 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     3961 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     4021 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     4081 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct
     4141 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     4201 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     4261 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     4321 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     4381 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     4441 cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag
     4501 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     4561 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     4621 cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc
     4681 ttgatcagat cttgatcccc tgcgccatca ga
//
