LOCUS       sequence-213198-        4658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Met-tRNA formyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4658)
  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 4658)
  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: B7-4
FEATURES             Location/Qualifiers
     source          1..4658
                     /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             1235..1252
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1326..1344)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1340..1387
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1996..2946)
                     /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      2994..3216
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3708..3802)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3833..4627)
                     /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(3944..3963)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4554..4573
                     /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 attcagaatc
      301 actacgtatt atttttgcgg gtacacctga ctttgcagcg cgtcatctcg acgcgctgtt
      361 gtcttctggt cataacgtcg ttggcgtgtt cacccagcca gaccgaccgg caggacgcgg
      421 taaaaaactg atgcccagcc cggttaaagt tctggctgag gaaaaaggtc tgcccgtttt
      481 tcaacctgtt tccctgcgtc cacaagaaaa ccagcaactg gtcgccgaac tgcaggctga
      541 tgttatggtc gtcgtcgcct atggtttaat tctgccgaaa gcagtgctgg agatgccgcg
      601 tcttggctgt atcaacgttc atggttcact gctgccacgc tggcgcggtg ctgcaccaat
      661 ccaacgctca ctatgggcgg gtgatgcaga aactggtgtg accattatgc aaatggatgt
      721 cggtttagac accggtgata tgctctataa gctctcctgc ccgattactg cagaagatac
      781 cagtggtacg ctgtacgaca agctggcaga gcttggccca caagggctta tcaccacgtt
      841 gaaacaactg gcagacggca cggcgaaacc agaagttcag gacgaaactc ttgtcactta
      901 cgccgagaag ttgagtaaag aagaagcgcg tattgactgg tcactttcgg cagcacagct
      961 tgaacgctgc attcgcgctt tcaatccatg gccaatgagc tggctggaaa ttgaaggaca
     1021 gccggttaaa gtctggaaag catcggtcat tgatacggca accaacgctg caccaggaac
     1081 gatccttgaa gccaacaaac aaggcattca ggttgcgact ggtgatggca tcctgaacct
     1141 gctctcgtta caacctgcgg gtaagaaagc gatgagcgcg caagacctcc tgaactctcg
     1201 tcgggaatgg tttgttccgg gcaaccgtct ggtccatcat catcaccatc attaaagcag
     1261 aggccatatg ctcgaggatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1321 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1381 ttttttgctg aaaggaggaa ctatatccgg atatcccgca agaggcccgg cagtaccggc
     1441 ataaccaagc ctatgcctac agcatccagg gcctagggta cgggttttgc tgcccgcaaa
     1501 cgggctgttc tggtgttgct agtttgttat cagaatcgca gatccggctt cagccggttt
     1561 gccggctgaa agcgctattt cttccagaat tgccatgatt ttttccccac gggaggcgtc
     1621 actggctccc gtgttgtcgg cagctttgat tcgataagca gcatcgcctg tttcaggctg
     1681 tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt gttcaatttc atgttctagt
     1741 tgctttgttt tactggtttc acctgttcta ttaggtgtta catgctgttc atctgttaca
     1801 ttgtcgatct gttcatggtg aacagctttg aatgcaccaa aaactcgtaa aagctctgat
     1861 gtatctatct tttttacacc gttttcatct gtgcatatgg acagttttcc ctttgatatg
     1921 taacggtgaa cagttgttct acttttgttt gttagtcttg atgcttcact gatagataca
     1981 agagccataa gaacctcaga tccttccgta tttagccagt atgttctcta gtgtggttcg
     2041 ttgtttttgc gtgagccatg agaacgaacc attgagatca tacttacttt gcatgtcact
     2101 caaaaatttt gcctcaaaac tggtgagctg aatttttgca gttaaagcat cgtgtagtgt
     2161 ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc
     2221 attcattttt atctggttgt tctcaagttc ggttacgaga tccatttgtc tatctagttc
     2281 aacttggaaa atcaacgtat cagtcgggcg gcctcgctta tcaaccacca atttcatatt
     2341 gctgtaagtg tttaaatctt tacttattgg tttcaaaacc cattggttaa gccttttaaa
     2401 ctcatggtag ttattttcaa gcattaacat gaacttaaat tcatcaaggc taatctctat
     2461 atttgccttg tgagttttct tttgtgttag ttcttttaat aaccactcat aaatcctcat
     2521 agagtatttg ttttcaaaag acttaacatg ttccagatta tattttatga atttttttaa
     2581 ctggaaaaga taaggcaata tctcttcact aaaaactaat tctaattttt cgcttgagaa
     2641 cttggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
     2701 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
     2761 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
     2821 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
     2881 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
     2941 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
     3001 tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat tctgctagac
     3061 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
     3121 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
     3181 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
     3241 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
     3301 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
     3361 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
     3421 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
     3481 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
     3541 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
     3601 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     3661 gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata
     3721 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     3781 actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga
     3841 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     3901 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     3961 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     4021 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     4081 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     4141 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     4201 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     4261 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     4321 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     4381 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     4441 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     4501 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     4561 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     4621 caatcatgcg aaacgatcct catcctgtct cttgatca
//
