LOCUS       sequence-213287-        5642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Thr-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5642)
  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 5642)
  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: B2-2
FEATURES             Location/Qualifiers
     source          1..5642
                     /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(2310..2328)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2324..2371
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2980..3930)
                     /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      3978..4200
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4692..4786)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4817..5611)
                     /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(4928..4947)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5538..5557
                     /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 cctgttataa ctcttcctga tggcagccaa cgccattacg atcacgctgt
      361 aagccccatg gatgttgcgc tggacattgg tccaggtctg gcgaaagcct gtatcgcagg
      421 gcgcgttaat ggcgaactgg ttgatgcttg cgatctgatt gaaaacgacg cacaactgtc
      481 gatcattacc gccaaagacg aagaaggtct ggagatcatt cgtcactcct gtgcgcacct
      541 gttagggcac gcgattaaac aactttggcc gcataccaaa atggcaatcg gcccggttat
      601 tgacaacggt ttttattacg acgttgatct tgaccgcacg ttaacccagg aagatgtcga
      661 agcactcgag aagcggatgc atgagcttgc tgagaaaaac tacgacgtca ttaagaagaa
      721 agtcagctgg cacgaagcgc gtgaaacttt cgccaaccgt ggggagagct acaaagtctc
      781 cattcttgac gaaaacatcg cccatgatga caagccaggt ctgtacttcc atgaagaata
      841 tgtcgatatg tgccgcggtc cgcacgtacc gaacatgcgt ttctgccatc atttcaaact
      901 aatgaaaacg gcaggggctt actggcgtgg cgacagcaac aacaaaatgt tgcaacgtat
      961 ttacggtacg gcgtgggcag acaaaaaagc acttaacgct tacctgcagc gcctggaaga
     1021 agccgcgaaa cgcgaccacc gtaaaatcgg taaacagctc gacctgtacc atatgcagga
     1081 agaagcgccg ggtatggtat tctggcacaa cgacggctgg accatcttcc gtgaactgga
     1141 agtgtttgtt cgttctaaac tgaaagagta ccagtatcag gaagttaaag gtccgttcat
     1201 gatggaccgt gtcctgtggg aaaaaaccgg tcactgggac aactacaaag atgcaatgtt
     1261 caccacatct tctgagaacc gtgaatactg cattaagccg atgaactgcc cgggtcacgt
     1321 acaaattttc aaccaggggc tgaagtctta tcgcgatctg ccgctgcgta tggccgagtt
     1381 tggtagctgc caccgtaacg agccgtcagg ttcgctgcat ggcctgatgc gcgtgcgtgg
     1441 atttacccag gatgacgcgc atatcttctg tactgaagaa caaattcgcg atgaagttaa
     1501 cggatgtatc cgtttagtct atgatatgta cagcactttt ggcttcgaga agatcgtcgt
     1561 caaactctcc actcgtcctg aaaaacgtat tggcagcgac gaaatgtggg atcgtgctga
     1621 ggcggacctg gcggttgcgc tggaagaaaa caacatcccg tttgaatatc aactgggtga
     1681 aggcgctttc tacggtccga aaattgaatt taccctgtat gactgcctcg atcgtgcatg
     1741 gcagtgcggt acagtacagc tggacttctc tttgccgtct cgtctgagcg cttcttatgt
     1801 aggcgaagac aatgaacgta aagtaccggt aatgattcac cgcgcaattc tggggtcgat
     1861 ggaacgtttc atcggtatcc tgaccgaaga gttcgctggt ttcttcccga cctggcttgc
     1921 gccggttcag gttgttatca tgaatattac cgattcacag tctgaatacg ttaacgaatt
     1981 gacgcaaaaa ctatcaaatg cgggcattcg tgttaaagca gacttgagaa atgagaagat
     2041 tggctttaaa atccgcgagc acactttgcg tcgcgtccca tatatgctgg tctgtggtga
     2101 taaagaggtg gaatcaggca aagttgccgt tcgcacccgc cgtggtaaag acctgggaag
     2161 catggacgta aatgaagtga tcgagaagct gcaacaagag attcgcagcc gcagtcttaa
     2221 acaattggag gaatattaaa gcagcggcca tatgctcgag gatccggctg ctaacaaagc
     2281 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2341 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
     2401 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggcctag
     2461 ggtacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat
     2521 cgcagatccg gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat
     2581 gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata
     2641 agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc
     2701 aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt
     2761 gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca
     2821 ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat
     2881 atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt
     2941 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     3001 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     3061 atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     3121 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt
     3181 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     3241 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     3301 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     3361 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     3421 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     3481 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     3541 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     3601 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     3661 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     3721 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     3781 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     3841 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     3901 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     3961 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     4021 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     4081 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     4141 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     4201 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     4261 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     4321 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     4381 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     4441 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     4501 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     4561 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     4621 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc
     4681 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     4741 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg
     4801 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4861 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4921 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4981 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     5041 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     5101 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     5161 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     5221 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     5281 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     5341 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     5401 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     5461 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     5521 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     5581 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     5641 ca
//
