LOCUS       sequence-213285-        5533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Asp-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5533)
  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 5533)
  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: B4-2
FEATURES             Location/Qualifiers
     source          1..5533
                     /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             2060..2077
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2201..2219)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2215..2262
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2871..3821)
                     /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      3869..4091
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4583..4677)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4708..5502)
                     /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(4819..4838)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5429..5448
                     /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 atcgtacaga
      301 atattgtgga cagctccgtt tgtcccacgt ggggcagcag gtgactctgt gtggttgggt
      361 caaccgtcgt cgtgatcttg gtagcctgat cttcatcgat atgcgcgacc gcgaaggtat
      421 cgtgcaggta tttttcgatc cggatcgtgc ggacgcgtta aagctggcct ctgaactgcg
      481 taatgagttc tgcattcagg tcacgggcac cgtacgtgcg cgtgacgaaa aaaatattaa
      541 ccgcgatatg gcgaccggcg aaatcgaagt gctggcgtcc tcgctgacta tcatcaaccg
      601 cgcagatgtt ctgccgcttg actctaacca cgtcaacacc gaagaagcgc gtctgaaata
      661 ccgctacctc gacctgcgtc gtccggaaat ggctcagcgc ctgaaaaccc gcgctaaaat
      721 caccagcctg gtgcgccgtt ttatggatga ccacggcttc ctcgacatcg aaactccgat
      781 gctgaccaaa gccacgccgg aaggcgcgcg tgactacctg gtgccttctc gtgtgcacaa
      841 aggtaaattc tacgcactgc cgcaatcccc gcagttgttc aaacagctgc tgatgatgtc
      901 cggttttgac cgttactatc agatcgttaa atgcttccgt gacgaagacc tgcgtgctga
      961 ccgtcagcct gaatttactc agatcgatgt ggaaacttct ttcatgaccg cgccgcaagt
     1021 gcgtgaagtg atggaagcgc tggtgcgtca tctgtggctg gaagtgaagg gtgtggatct
     1081 gggcgatttc ccggtaatga cctttgcgga agcagaacgc cgttatggtt ctgataaacc
     1141 ggatctgcgt aacccgatgg aactgactga cgttgctgat ctgctgaaat ctgttgagtt
     1201 tgctgtattt gcaggtccgg cgaacgatcc gaaaggtcgc gtagcggctc tgcgcgttcc
     1261 gggcggcgca tcgctgaccc gtaagcagat cgacgaatac ggtaacttcg ttaaaatcta
     1321 cggcgcgaaa ggtctggctt acatcaaagt taacgaacgc gcgaaaggtc tggaaggtat
     1381 caacagcccg gtagcgaagt tccttaatgc agaaatcatc gaagacatcc tggatcgtac
     1441 tgccgcgcaa gatggcgata tgattttctt cggtgccgac aacaagaaaa ttgttgccga
     1501 cgcgatgggt gcactgcgcc tgaaagtggg taaagacctt ggtctgaccg acgaaagcaa
     1561 atgggcaccg ctgtgggtta tcgacttccc gatgtttgaa gacgacggtg aaggcggcct
     1621 gacggcaatg caccatccgt tcacctcacc gaaagatatg acggctgcag aactgaaagc
     1681 tgcaccggaa aatgcggtgg cgaacgctta cgatatggtc atcaatggtt acgaagtggg
     1741 cggtggttca gtacgtatcc ataatggtga tatgcagcag acggtgtttg gtattctggg
     1801 tatcaacgaa gaggaacagc gcgagaaatt cggcttcctg ctcgacgctc tgaaatacgg
     1861 tactccgccg cacgcaggtc tggcattcgg tcttgaccgt ctgaccatgc tgctgaccgg
     1921 caccgacaat atccgtgacg ttatcgcctt cccgaaaacc acggcggcag cgtgtctgat
     1981 gactgaagca ccgagctttg ctaacccgac tgcactggct gagctgagca ttcaggttgt
     2041 gaagaaggct gagaataacc atcatcatca ccatcattaa agcgttgtgc ctgtagccga
     2101 cgtactccgc gtaacgccag cttctattaa agcagcggcc atatgctcga ggatccggct
     2161 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2221 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     2281 tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     2341 ccagggccta gggtacgggt tttgctgccc gcaaacgggc tgttctggtg ttgctagttt
     2401 gttatcagaa tcgcagatcc ggcttcagcc ggtttgccgg ctgaaagcgc tatttcttcc
     2461 agaattgcca tgattttttc cccacgggag gcgtcactgg ctcccgtgtt gtcggcagct
     2521 ttgattcgat aagcagcatc gcctgtttca ggctgtctat gtgtgactgt tgagctgtaa
     2581 caagttgtct caggtgttca atttcatgtt ctagttgctt tgttttactg gtttcacctg
     2641 ttctattagg tgttacatgc tgttcatctg ttacattgtc gatctgttca tggtgaacag
     2701 ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
     2761 catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt gttctacttt
     2821 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
     2881 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
     2941 gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc aaaactggtg
     3001 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt atgtaggtag
     3061 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
     3121 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
     3181 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
     3241 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
     3301 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
     3361 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
     3421 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
     3481 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     3541 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     3601 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     3661 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     3721 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     3781 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     3841 tgattttaat cactatacca attgagatgg gctagtcaat gataattact agtccttttc
     3901 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     3961 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     4021 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     4081 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     4141 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     4201 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     4261 ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca
     4321 ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt
     4381 cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc
     4441 tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca
     4501 ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg
     4561 tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     4621 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     4681 gcgagctctc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     4741 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     4801 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     4861 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     4921 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt
     4981 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     5041 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     5101 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     5161 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     5221 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     5281 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc
     5341 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     5401 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     5461 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     5521 tgtctcttga tca
//
