LOCUS       sequence-231791-        4562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  translational elongation factor Ts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4562)
  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 4562)
  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: B1-5
FEATURES             Location/Qualifiers
     source          1..4562
                     /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             940..957
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1031..1049)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1045..1092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1701..2651)
                     /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      2699..2921
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3413..3507)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3538..4332)
                     /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(3649..3668)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4259..4278
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgagag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg catgctgaaa ttaccgcatc cctggtaaaa
      121 gagctgcgtg agcgtactgg cgcaggcatg atggattgca aaaaagcact gactgaagct
      181 aacggcgaca tcgagctggc aatcgaaaac atgcgtaagt ccggtgctat taaagcagcg
      241 aaaaaagcag gcaacgttgc tgctgacggc gtgatcaaaa ccaaaatcga cggcaactac
      301 ggcatcattc tggaagttaa ctgccagact gacttcgttg caaaagacgc tggtttccag
      361 gcgttcgcag acaaagttct ggacgcagct gttgctggca aaatcactga cgttgaagtt
      421 ctgaaagcac agttcgaaga agaacgtgtt gcgctggtag cgaaaattgg tgaaaacatc
      481 aacattcgcc gcgttgctgc gctggaaggc gacgttctgg gttcttatca gcacggtgcg
      541 cgtatcggcg ttctggttgc tgctaaaggc gctgacgaag agctggttaa acacatcgct
      601 atgcacgttg ctgcaagcaa gccagaattc atcaaaccgg aagacgtatc cgctgaagtg
      661 gtagaaaaag aataccaggt acagctggat atcgcgatgc agtctggtaa gccgaaagaa
      721 atcgcagaga aaatggttga aggccgcatg aagaaattca ccggcgaagt ttctctgacc
      781 ggtcagccgt tcgttatgga accaagcaaa actgttggtc agctgctgaa agagcataac
      841 gctgaagtga ctggcttcat ccgcttcgaa gtgggtgaag gcatcgagaa agttgagact
      901 gactttgcag cagaagttgc tgcgatgtcc aagcagtctc atcatcatca ccatcattaa
      961 agcagcggcc atatgctcga ggatccggct gctaacaaag cccgaaagga agctgagttg
     1021 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1081 aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta
     1141 ccggcataac caagcctatg cctacagcat ccagggccta gggtacgggt tttgctgccc
     1201 gcaaacgggc tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc
     1261 ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag
     1321 gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca
     1381 ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt
     1441 ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg
     1501 ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct
     1561 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg
     1621 atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag
     1681 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg
     1741 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg
     1801 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt
     1861 agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg
     1921 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct
     1981 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc
     2041 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt
     2101 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc
     2161 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc
     2221 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt
     2281 tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt
     2341 gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg
     2401 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt
     2461 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct
     2521 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg
     2581 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact
     2641 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg
     2701 gctagtcaat gataattact agtccttttc ctttgagttg tgggtatctg taaattctgc
     2761 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct
     2821 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa
     2881 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg
     2941 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac
     3001 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc
     3061 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     3121 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     3181 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     3241 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     3301 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     3361 aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac
     3421 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     3481 tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga gtcccgctca
     3541 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     3601 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     3661 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     3721 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     3781 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     3841 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     3901 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     3961 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     4021 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     4081 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     4141 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     4201 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     4261 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     4321 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg
     4381 ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt
     4441 accagagggc gccccagctg gcaattccga cgtctaagaa accattatta tcatgacatt
     4501 aacctataaa aataggcgga tcacgaggcc ctttcgtctt cacctcgatg atcccgcgaa
     4561 at
//
