LOCUS       sequence-267514-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gln-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  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 5378)
  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: B7-2
FEATURES             Location/Qualifiers
     source          1..5378
                     /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             91..108
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1847..1865)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1861..1908
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2517..3467)
                     /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      3515..3737
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4229..4323)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4354..5148)
                     /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(4465..4484)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5075..5094
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgagag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg catcatcatc accatcattt aagtgaggca
      121 gaagcccgcc cgactaactt tatccgtcag atcatcgatg aagatctggc cagtggtaag
      181 cacaccacag tacacacccg tttcccgccg gagccgaatg gctatctgca tattggccat
      241 gcgaaatcta tctgcctgaa cttcgggatc gcccaggact ataaaggcca gtgcaacctg
      301 cgtttcgacg acactaaccc ggtaaaagaa gatatcgagt atgttgagtc gatcaaaaac
      361 gacgtagagt ggttaggttt tcactggtct ggtaacgtcc gttactcctc cgattatttt
      421 gatcagctcc acgcctatgc gatcgaactg atcaataaag gcctggcgta cgttgatgaa
      481 ctgacgccgg aacagatccg cgaataccgc ggcaccctga cgcaaccggg taaaaacagc
      541 ccgtaccgcg accgcagcgt tgaagagaac ctggcgctgt tcgaaaaaat gcgtgccggt
      601 ggttttgaag aaggtaaagc ctgcctgcgt gcgaaaatcg acatggcttc accgtttatc
      661 gtgatgcgcg atccggtgct gtaccgtatt aaatttgctg aacaccacca gactggcaac
      721 aagtggtgca tctacccgat gtacgacttc acccactgca tcagcgatgc cctggaaggt
      781 attacgcact ctctgtgtac gcttgagttc caggacaacc gtcgtctgta cgactgggta
      841 ctggacaaca tcacgattcc tgttcacccg cgccagtatg agttctcgcg cctgaatctg
      901 gaatacaccg tgatgtccaa gcgtaagttg aacctgctgg tgaccgacaa gcacgttgaa
      961 ggctgggatg acccgcgtat gccgaccatt tccggtctgc gtcgtcgtgg ttacactgcg
     1021 gcttctattc gtgagttctg caaacgcatc ggcgtgacca agcaggacaa caccattgag
     1081 atggcgtcgc tggaatcctg catccgtgaa gatctcaacg aaaatgcgcc gcgcgcaatg
     1141 gcggttatcg atccggtgaa actggttatc gaaaactatc agggcgaagg cgaaatggtt
     1201 accatgccga accatccgaa caaaccggaa atgggcagcc gtcaggtgcc gtttagcggt
     1261 gagatttgga ttgatcgcgc cgatttccgc gaagaagcta acaagcagta caaacgtctg
     1321 gtgctgggta aagaagtgcg tctgcgtaat gcttatgtga ttaaggcaga acgcgtcgag
     1381 aaagatgccg aaggtaatat caccaccatc ttctgtactt atgacgccga taccttaagc
     1441 aaagatccgg cagatggtcg taaagtcaaa ggtgttattc actgggtgag cgcggcacat
     1501 gcgctgccgg ttgaaatccg tttgtatgac cgtctgttca gcgtgcctaa cccaggtgct
     1561 gcggatgatt tcctgtcggt gattaacccg gaatcgctgg tgatcaaaca gggctttgct
     1621 gaaccgtcgc tgaaagatgc ggttgcgggt aaagcattcc agtttgagcg tgaaggttac
     1681 ttctgcctcg atagccgcca ttctacggcg gaaaaacctg tatttaaccg caccgttggg
     1741 ctgcgtgata cctgggcgaa agtaggcgag tattaaagca gaggccatat gctcgaggat
     1801 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1861 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     1921 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     1981 cagcatccag ggcctagggt acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc
     2041 tagtttgtta tcagaatcgc agatccggct tcagccggtt tgccggctga aagcgctatt
     2101 tcttccagaa ttgccatgat tttttcccca cgggaggcgt cactggctcc cgtgttgtcg
     2161 gcagctttga ttcgataagc agcatcgcct gtttcaggct gtctatgtgt gactgttgag
     2221 ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt ttactggttt
     2281 cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc tgttcatggt
     2341 gaacagcttt gaatgcacca aaaactcgta aaagctctga tgtatctatc ttttttacac
     2401 cgttttcatc tgtgcatatg gacagttttc cctttgatat gtaacggtga acagttgttc
     2461 tacttttgtt tgttagtctt gatgcttcac tgatagatac aagagccata agaacctcag
     2521 atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat
     2581 gagaacgaac cattgagatc atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa
     2641 ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttatgt
     2701 aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg
     2761 ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta
     2821 tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct
     2881 ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca
     2941 agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc
     3001 ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa
     3061 gacttaacat gttccagatt atattttatg aattttttta actggaaaag ataaggcaat
     3121 atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac
     3181 tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc
     3241 tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga
     3301 gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg
     3361 gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag
     3421 cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg
     3481 tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc
     3541 cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta
     3601 aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt
     3661 caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc
     3721 agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa
     3781 acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc
     3841 gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg
     3901 ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa
     3961 atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa
     4021 gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact
     4081 ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc
     4141 gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaggct
     4201 tacccgtctt actgtcccta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     4261 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     4321 gtccaagcga gctctcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     4381 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     4441 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     4501 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     4561 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     4621 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     4681 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     4741 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     4801 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     4861 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     4921 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     4981 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     5041 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     5101 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     5161 tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     5221 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     5281 ttccgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcggatcac
     5341 gaggcccttt cgtcttcacc tcgatgatcc cgcgaaat
//
