LOCUS       sequence-213290-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  translational elongation factor 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  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 5510)
  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: B3-2
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     92..111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        92..110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             150..172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1979..1996
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2070..2088)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2084..2131
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2740..3690)
                     /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      3738..3960
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4452..4546)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4577..5371)
                     /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(4688..4707)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5298..5317
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtcttc acctcgatga tcccgcgaaa ttaatacgac tcactatagg ggaattgaga
      121 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat
      181 gcataagaat atacgtaact tttcgatcat agctcacatt gaccacggta aatcgacgct
      241 gtctgaccgt attatccaga tctgcggtgg cctgtctgac cgtgaaatgg aggcgcaggt
      301 tctcgattcc atggatcttg agcgtgagcg tggcattacc atcaaagcgc aaagcgtgac
      361 gctggactac aaagcgtctg acggcgaaac ctatcagctt aactttatcg acaccccggg
      421 ccacgtagac ttctcctatg aagtttcccg ttcgctggct gcctgtgaag gtgcattgct
      481 ggtggtcgac gccgggcagg gcgtagaagc gcaaaccctg gcaaactgct acaccgccat
      541 ggaaatggat ctcgaagttg tgccggtact gaacaagatt gacctgccgg cagccgatcc
      601 tgaacgcgtg gcggaagaaa ttgaagatat cgtcggcatc gacgccaccg acgcggtgcg
      661 ctgttcagcg aaaaccggcg ttggtgtgca ggacgttctc gaacgtctgg tgcgcgacat
      721 tccgccgccg gaaggcgatc cggaaggccc gttgcaggca ctaattatcg actcatggtt
      781 cgacaactac ctgggcgttg tttcacttat ccgtattaaa aacggcaccc tgcgtaaggg
      841 cgacaaagtg aaagtcatga gtaccgggca gacctataac gccgaccgtc tgggcatctt
      901 cacgccgaaa caggttgacc gcactgaact gaaatgtggc gaagtaggct ggctcgtatg
      961 tgcgattaaa gatatccacg gcgctccagt cggcgatacc ttaacgctgg cgcgtaatcc
     1021 ggcagaaaag gcgctgcctg gctttaagaa agtcaaaccg caggtatacg ccggtctgtt
     1081 cccggtaagt tccgacgact atgaagcctt ccgtgacgcg ctgggtaaac tcagcctgaa
     1141 cgatgcctca ctgttctatg agccggaaag ctccagcgcg ctgggctttg gtttccgctg
     1201 cggcttcctc ggcctgctgc acatggagat catccaggaa cgtctggaac gtgaatacga
     1261 tctggatctg atcaccactg cgccgaccgt agtgtatgaa gttgaaacca cgtcaagaga
     1321 agttatctac gtcgacagcc catccaagct gcctgcggta aataacatct acgaactgcg
     1381 cgagccgatt gcagagtgtc acatgctgct gccgcaggca tatctcggca acgttattac
     1441 gttgtgcgta gaaaaacgcg gcgtgcagac caatatggtt taccacggta atcaggtggc
     1501 gctgacgtac gagatcccga tggcggaagt ggtgctcgac ttcttcgatc gcctgaaatc
     1561 tacctcgcgt ggttatgcgt ctctggatta caacttcaag cgcttccagg cgtccgacat
     1621 ggtacgtgta gacgtattaa tcaacggtga acgtgttgat gcgctggcgt tgatcaccca
     1681 ccgtgataat tcgcaaaacc gcggtcgcga gttggtggag aagatgaaag atctgatccc
     1741 acgccagcag tttgatatcg ccattcaggc agcgattggt acgcacatca ttgcgcgatc
     1801 caccgtgaaa cagctgcgta aaaacgtact ggctaaatgt tatggcggcg atatcagccg
     1861 taagaaaaag ctgctgcaga agcagaaaga aggtaagaaa cgcatgaagc agatcggtaa
     1921 cgtcgagctg ccgcaggaag cgttcctcgc cattctgcac gtcggcaaag acaacaaaca
     1981 tcatcatcac catcattaaa gcagaggcca tatgctcgag gatccggctg ctaacaaagc
     2041 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2101 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
     2161 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggcctag
     2221 ggtacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg ttatcagaat
     2281 cgcagatccg gcttcagccg gtttgccggc tgaaagcgct atttcttcca gaattgccat
     2341 gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt tgattcgata
     2401 agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac aagttgtctc
     2461 aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt tctattaggt
     2521 gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc tttgaatgca
     2581 ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc atctgtgcat
     2641 atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt gtttgttagt
     2701 cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc cgtatttagc
     2761 cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg aaccattgag
     2821 atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga gctgaatttt
     2881 tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg aatctgatgt
     2941 aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa gttcggttac
     3001 gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg ggcggcctcg
     3061 cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta ttggtttcaa
     3121 aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta acatgaactt
     3181 aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg ttagttcttt
     3241 taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa catgttccag
     3301 attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt cactaaaaac
     3361 taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa tctcaaagcc
     3421 tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg ttgctttagc
     3481 taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt ggttataagt
     3541 gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga gtagtgccac
     3601 acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa tcgctagttc
     3661 atttgctttg aaaacaacta attcagacat acatctcaat tggtctaggt gattttaatc
     3721 actataccaa ttgagatggg ctagtcaatg ataattacta gtccttttcc tttgagttgt
     3781 gggtatctgt aaattctgct agacctttgc tggaaaactt gtaaattctg ctagaccctc
     3841 tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat attcaagtgg ttataattta
     3901 tagaataaag aaagaataaa aaaagataaa aagaatagat cccagccctg tgtataactc
     3961 actactttag tcagttccgc agtattacaa aaggatgtcg caaacgctgt ttgctcctct
     4021 acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc ctcgcaagct cgggcaaatc
     4081 gctgaatatt ccttttgtct ccgaccatca ggcacctgag tcgctgtctt tttcgtgaca
     4141 ttcagttcgc tgcgctcacg gctctggcag tgaatggggg taaatggcac tacaggcgcc
     4201 ttttatggat tcatgcaagg aaactaccca taatacaaga aaagcccgtc acgggcttct
     4261 cagggcgttt tatggcgggt ctgctatgtg gtgctatctg actttttgct gttcagcagt
     4321 tcctgccctc tgattttcca gtctgaccac ttcggattat cccgtgacag gtcattcaga
     4381 ctggctaatg cacccagtaa ggcagcggta tcatcaacag gcttacccgt cttactgtcc
     4441 ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa
     4501 tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctctcg
     4561 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4621 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4681 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4741 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4801 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     4861 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4921 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4981 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     5041 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     5101 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     5161 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     5221 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     5281 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     5341 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     5401 cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     5461 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccgac
//
