LOCUS       sequence-213281-        4256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  translational initiation factor 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4256)
  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 4256)
  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: B5-5
FEATURES             Location/Qualifiers
     source          1..4256
                     /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             290..307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             800..811
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(924..942)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      938..985
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1594..2544)
                     /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      2592..2814
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3306..3400)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3431..4225)
                     /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(3542..3561)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4152..4171
                     /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 atcatcatca
      301 ccatcattta aaaggcggaa aacgagttca aacggcgcgc cctaaccgta tcaatggcga
      361 aattcgcgcc caggaagttc gcttaacagg tctggaaggc gagcagcttg gtattgtgag
      421 tctgagagaa gctctggaga aagcagaaga agccggagta gacttagtcg agatcagccc
      481 taacgccgag ccgccggttt gtcgtataat ggattacggc aaattcctct atgaaaagag
      541 caagtcttct aaggaacaga agaaaaagca aaaagttatc caggttaagg aaattaaatt
      601 ccgtcctggt acagatgaag gcgactatca ggtaaaactc cgcagcctga ttcgctttct
      661 cgaagagggt gataaagcca aaatcacgct gcgtttccgc ggtcgtgaga tggcgcacca
      721 gcaaatcggt atggaagtgc ttaatcgcgt gaaagacgat ttgcaagaac tggcagtggt
      781 cgaatccttc ccaacgaaga tcgaaggccg ccagatgatc atggtgctcg ctcctaagaa
      841 gaaacagtat taaagcagag gccatatgct cgaggatccg gctgctaaca aagcccgaaa
      901 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
      961 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat atcccgcaag
     1021 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggc ctagggtacg
     1081 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     1141 tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt
     1201 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc
     1261 atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt
     1321 tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca
     1381 tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa
     1441 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     1501 agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     1561 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     1621 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata
     1681 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     1741 taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt
     1801 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     1861 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     1921 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     1981 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     2041 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     2101 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     2161 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     2221 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac
     2281 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     2341 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     2401 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     2461 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     2521 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     2581 ccaattgaga tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat
     2641 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa
     2701 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     2761 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     2821 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     2881 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     2941 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     3001 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
     3061 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
     3121 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
     3181 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
     3241 aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg
     3301 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     3361 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct ctcgaacccc
     3421 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     3481 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     3541 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     3601 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     3661 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     3721 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     3781 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3841 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3901 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3961 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4021 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     4081 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     4141 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     4201 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatca
//
