LOCUS       sequence-213284-        5126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Glu-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5126)
  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 5126)
  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: B3-3
FEATURES             Location/Qualifiers
     source          1..5126
                     /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             1703..1720
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1794..1812)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1808..1855
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2464..3414)
                     /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      3462..3684
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4176..4270)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4301..5095)
                     /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(4412..4431)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5022..5041
                     /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 ataaaatcaa
      301 aactcgcttc gcgccaagcc caacaggcta tctgcacgtt ggcggcgcgc gtactgctct
      361 ttactcctgg ctttttgcac gtaaccacgg cggtgagttc gtgctgcgta ttgaagacac
      421 cgatcttgag cgttccacgc cggaagctat cgaagccatt atggatggca tgaactggct
      481 gagcctggag tgggatgaag gtccgtacta ccagaccaaa cgttttgatc gctacaacgc
      541 ggtgatcgat cagatgctgg aagagggcac tgcttataaa tgctattgct ctaaagagcg
      601 cctggaagcg ctgcgcgaag agcaaatggc gaaaggtgag aagccgcgtt atgacggtcg
      661 ctgccgccac agccatgagc atcatgctga tgatgaaccg tgtgttgtac gttttgctaa
      721 cccgcaggaa ggttctgttg tttttgacga tcagatccgt ggtccgatcg agttcagcaa
      781 ccaggaactg gacgatctta ttatccgccg taccgatggt tccccaacct ataacttctg
      841 tgtggttgtc gatgactggg atatggaaat cacccacgtt atccgtggcg aagaccatat
      901 caacaacacg ccacgccaga tcaacattct taaggccctg aaagcgccgg tgccggttta
      961 cgcgcacgtt tctatgatca atggcgatga cggtaaaaaa ctgtccaaac gtcacggggc
     1021 agtcagcgta atgcagtatc gtgatgacgg ttatttgcca gaagcactgc tgaactatct
     1081 ggtgcgtctg ggctggtccc acggcgatca ggaaatcttc actcgtgaag agatgatcaa
     1141 atacttcact ttgaatgccg tcagcaaatc tgccagtgcg ttcaacaccg acaagctgct
     1201 gtggctgaac catcactaca ttaacgcgct gccgccggag tatgttgcta ctcacttaca
     1261 gtggcacatt gagcaggaaa atatcgatac ccgtaacggc ccgcagctgg ctgatctggt
     1321 gaaactgctg ggcgaacgct gcaagacgct gaaagagatg gcacagagct gccgttattt
     1381 ctacgaagat tttgctgagt tcgatgccga cgccgcgaaa aaacatctgc gtccggtagc
     1441 gcgtcagccg ctggaagtgg ttcgtgacaa actggccgcg attactgact ggaccgctga
     1501 aaacgttcat cacgctattc aggcgacggc ggatgagctg gaagtgggta tgggtaaagt
     1561 tggtatgccg ctgcgtgtcg ccgtaaccgg tgcggggcag tctccagcac tggatgttac
     1621 cgttcacgca attggtaaga cccgcagtat cgagcgtatc aacaaagcgc tggattttat
     1681 tgctgaacgc gaaaatcagc agcatcatca tcaccatcat taaagcagag gccatatgct
     1741 cgaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1801 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1861 aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     1921 atgcctacag catccagggc ctagggtacg ggttttgctg cccgcaaacg ggctgttctg
     1981 gtgttgctag tttgttatca gaatcgcaga tccggcttca gccggtttgc cggctgaaag
     2041 cgctatttct tccagaattg ccatgatttt ttccccacgg gaggcgtcac tggctcccgt
     2101 gttgtcggca gctttgattc gataagcagc atcgcctgtt tcaggctgtc tatgtgtgac
     2161 tgttgagctg taacaagttg tctcaggtgt tcaatttcat gttctagttg ctttgtttta
     2221 ctggtttcac ctgttctatt aggtgttaca tgctgttcat ctgttacatt gtcgatctgt
     2281 tcatggtgaa cagctttgaa tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt
     2341 tttacaccgt tttcatctgt gcatatggac agttttccct ttgatatgta acggtgaaca
     2401 gttgttctac ttttgtttgt tagtcttgat gcttcactga tagatacaag agccataaga
     2461 acctcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt
     2521 gagccatgag aacgaaccat tgagatcata cttactttgc atgtcactca aaaattttgc
     2581 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc
     2641 gttatgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat
     2701 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat
     2761 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt
     2821 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt
     2881 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg
     2941 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt
     3001 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata
     3061 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt
     3121 tgtccactgg aaaatctcaa agcctttaac caaaggattc ctgatttcca cagttctcgt
     3181 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat
     3241 catctgagcg tattggttat aagtgaacga taccgtccgt tctttccttg tagggttttc
     3301 aatcgtgggg ttgagtagtg ccacacagca taaaattagc ttggtttcat gctccgttaa
     3361 gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca actaattcag acatacatct
     3421 caattggtct aggtgatttt aatcactata ccaattgaga tgggctagtc aatgataatt
     3481 actagtcctt ttcctttgag ttgtgggtat ctgtaaattc tgctagacct ttgctggaaa
     3541 acttgtaaat tctgctagac cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt
     3601 ttatattcaa gtggttataa tttatagaat aaagaaagaa taaaaaaaga taaaaagaat
     3661 agatcccagc cctgtgtata actcactact ttagtcagtt ccgcagtatt acaaaaggat
     3721 gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa aaccctaaag gcttaagtag
     3781 caccctcgca agctcgggca aatcgctgaa tattcctttt gtctccgacc atcaggcacc
     3841 tgagtcgctg tctttttcgt gacattcagt tcgctgcgct cacggctctg gcagtgaatg
     3901 ggggtaaatg gcactacagg cgccttttat ggattcatgc aaggaaacta cccataatac
     3961 aagaaaagcc cgtcacgggc ttctcagggc gttttatggc gggtctgcta tgtggtgcta
     4021 tctgactttt tgctgttcag cagttcctgc cctctgattt tccagtctga ccacttcgga
     4081 ttatcccgtg acaggtcatt cagactggct aatgcaccca gtaaggcagc ggtatcatca
     4141 acaggcttac ccgtcttact gtccctagtg cttggattct caccaataaa aaacgcccgg
     4201 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
     4261 aacaggagtc caagcgagct ctcgaacccc agagtcccgc tcagaagaac tcgtcaagaa
     4321 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     4381 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     4441 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     4501 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     4561 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     4621 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     4681 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     4741 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     4801 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     4861 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     4921 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     4981 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     5041 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     5101 acgatcctca tcctgtctct tgatca
//
