LOCUS       sequence-213280-        5096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cys-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5096)
  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 5096)
  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: B5-3
FEATURES             Location/Qualifiers
     source          1..5096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1673..1690
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1764..1782)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1778..1825
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2434..3384)
                     /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      3432..3654
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4146..4240)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4271..5065)
                     /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(4382..4401)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4992..5011
                     /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 aggcggatca cgaggccctt tcgtcttcac
      181 ctcgatgatc ccgcgaaatt aatacgactc actatagggg aattgagagc ggataacaat
      241 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgc atctaaaaat
      301 cttcaatact ctgacacgcc aaaaagagga atttaagcct attcacgccg gggaagtcgg
      361 catgtacgtg tgtggaatca ccgtttacga tctctgtcat atcggtcacg ggcgtacctt
      421 tgttgctttt gacgtggttg cgcgctatct gcgtttcctc ggctataaac tgaagtatgt
      481 gcgcaacatt accgatatcg acgacaaaat catcaaacgc gccaatgaaa atggcgaaag
      541 ctttgtggcg atggtggatc gcatgatcgc cgaaatgcac aaagattttg atgctttgaa
      601 cattctgcgc ccggatatgg agccgcgcgc gacgcaccat atcgcagaaa ttattgaact
      661 cactgaacaa ctgatcgcca aaggtcacgc ttatgtggcg gacaacggcg acgtgatgtt
      721 cgacgtcccg accgatccaa cttatggcgt gctgtcgcgt caggatctcg accagctgca
      781 ggcaggcgcg cgcgttgacg tggtcgacga caaacgcaac ccaatggact tcgttctgtg
      841 gaagatgtcg aaagagggcg aaccgagctg gccgtctccg tggggcgcgg gtcgtcctgg
      901 ctggcacatt gaatgttcgg caatgaactg caagcagctg ggtaaccact ttgatatcca
      961 cggcggcggt tcagacctga tgttcccgca ccacgaaaac gaaatcgcgc agtccacctg
     1021 tgcccatgat ggtcagtatg tgaactactg gatgcactcg gggatggtga tggttgaccg
     1081 cgagaagatg tccaaatcgc tgggtaactt ctttaccgtg cgcgatgtgc tgaaatacta
     1141 cgacgcggaa accgtgcgtt acttcctgat gtcgggccac tatcgcagcc agttgaacta
     1201 cagcgaagag aacctgaagc aggcgcgtgc ggcgctggag cgtctctaca ctgcgctgcg
     1261 cggcacagat aaaaccgttg cgcctgccgg tggcgaagcg tttgaagcgc gctttattga
     1321 agcgatggac gacgatttca acaccccgga agcctattcc gtactgtttg atatggcgcg
     1381 tgaagtaaac cgtctgaaag cagaagatat ggcagcggcg aatgcaatgg catctcacct
     1441 gcgtaaactt tccgctgtat tgggcctgct ggagcaagaa ccggaagcgt tcctgcaaag
     1501 cggcgcgcag gcagacgaca gcgaagtggc tgagattgaa gcgttaattc aacagcgtct
     1561 ggatgcccgt aaagcgaaag actgggcggc ggcggatgcg gcgcgtgatc gtcttaacga
     1621 gatggggatc gtgctggaag atggcccgca agggaccacc tggcgtcgta agcatcatca
     1681 tcaccatcat taaagcagcg gccatatgct cgaggatccg gctgctaaca aagcccgaaa
     1741 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1801 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag
     1861 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggc ctagggtacg
     1921 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     1981 tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt
     2041 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc
     2101 atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt
     2161 tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca
     2221 tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa
     2281 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     2341 agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     2401 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     2461 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata
     2521 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     2581 taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt
     2641 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     2701 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     2761 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     2821 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     2881 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     2941 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     3001 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     3061 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac
     3121 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     3181 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     3241 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     3301 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     3361 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     3421 ccaattgaga tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat
     3481 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa
     3541 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     3601 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     3661 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     3721 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     3781 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     3841 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
     3901 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
     3961 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
     4021 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
     4081 aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg
     4141 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     4201 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct ctcgaacccc
     4261 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4321 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4381 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     4441 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     4501 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     4561 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4621 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     4681 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     4741 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     4801 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4861 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     4921 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     4981 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     5041 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatca
//
