LOCUS       sequence-330056-        4418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses heterlogous ribosomal proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4418)
  AUTHORS   Kolber NS, Fattal R, Bratulic S, Carver GD, Badran AH
  TITLE     Orthogonal translation enables heterologous ribosome engineering in 
            E. coli.
  JOURNAL   Nat Commun. 2021 Jan 26;12(1):599. doi: 10.1038/s41467-020-20759-z.
  PUBMED    33500394
REFERENCE   2  (bases 1 to 4418)
  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"; journalName: "Nat 
            Commun."; date: "2021-01-26"; volume: "12(1):599. doi"; pages: " 
            10.1038/s41467-020-20759-z"
COMMENT     Sequence Label: pNK083b
FEATURES             Location/Qualifiers
     source          1..4418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      184..406
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(968..1846)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1873..2157
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2080..2099
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      complement(2783..2814)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2830..3489)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(join(3604..4418,1..136))
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta
       61 gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa
      121 caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga
      181 gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt atctgtaaat
      241 tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta
      301 gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag
      361 aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag
      421 ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt
      481 aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt
      541 ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg
      601 ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat
      661 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
      721 gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat
      781 tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc
      841 cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtcaagag gacatccggt
      901 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggagacttag
      961 ggacccttta tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga
     1021 actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt
     1081 caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct
     1141 tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc
     1201 ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat
     1261 caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat
     1321 ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa
     1381 gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt
     1441 gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat
     1501 tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa
     1561 cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg
     1621 gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca
     1681 ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga
     1741 taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat
     1801 taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc
     1861 caccattcag agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
     1921 tttactggct cttctcgcta acccaaccgg taaccccgct tattaaaagc attctgtaac
     1981 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
     2041 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     2101 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     2161 cgttttacct gcaggtgcag ttattttcaa ggagtgcaag agtggcaaac tctgctcaag
     2221 ctaaaaaacg tgcgcgccaa aacgttaaag cgcgtaaaca caacgcaagc ttgcgttcta
     2281 tggttcgtac ttacatcaaa cgtactttaa gtgcaatcgc tggtggtgat tatgctgttg
     2341 ctacagaagc ttacaaaaaa gctgttcctg taatcgaccg catggctgat aaaggcatca
     2401 tccacaaaaa taaagctgct cgtcataaga gccgtttaaa tgctcaagtt aaagctttag
     2461 ctaactaatt gaagcaagaa ttataaggct tttatcatgg ttgttattcg tttagcacgt
     2521 ggcggcgcaa aaaaacgtcc attctatcaa atcgttgtga ctgatagccg caatgcacgt
     2581 gatggtcgtt tcatcgagcg tatcggtttc tttaacccaa ctgcacaagg tcaagcagaa
     2641 aaacttcgtt tagacgcaga ccgttttgct cactgggttt ctcaaggtgc tcaaccttct
     2701 gaacgtgttg cttctttggc tgctcaagct aaaaaagcta ctgcataatt attgtaactt
     2761 aattaacggc actcctcagc caagtcaaaa gcctccggtc ggaggctttt gactacatgc
     2821 ccatggcgtt tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     2881 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     2941 caccttgtcg ccttgcgtat aatatttgcc catagtgaaa acgggggcga agaagttgtc
     3001 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     3061 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     3121 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     3181 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     3241 caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc
     3301 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     3361 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagtaactg actgaaatgc
     3421 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     3481 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     3541 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccagg ccaaataggc
     3601 cgttcagatc cttccgtatt tagccagtat gttctctagt gtggttcgtt gtttttgcgt
     3661 gagccatgag aacgaaccat tgagatcatg cttactttgc atgtcactca aaaattttgc
     3721 ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc
     3781 gttacgtagg taggaatctg atgtaatggt tgttggtatt ttgtcaccat tcatttttat
     3841 ctggttgttc tcaagttcgg ttacgagatc catttgtcta tctagttcaa cttggaaaat
     3901 caacgtatca gtcgggcggc ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt
     3961 taaatcttta cttattggtt tcaaaaccca ttggttaagc cttttaaact catggtagtt
     4021 attttcaagc attaacatga acttaaattc atcaaggcta atctctatat ttgccttgtg
     4081 agttttcttt tgtgttagtt cttttaataa ccactcataa atcctcatag agtatttgtt
     4141 ttcaaaagac ttaacatgtt ccagattata ttttatgaat ttttttaact ggaaaagata
     4201 aggcaatatc tcttcactaa aaactaattc taatttttcg cttgagaact tggcatagtt
     4261 tgtccactgg aaaatcttaa agcctttaac caaaggattc ctgatttcca cagttctcgt
     4321 catcagctct ctggttgctt tagctaatac accataagca ttttccctac tgatgttcat
     4381 catctgagcg tattggttat aagtgaacga taccgtcc
//
