LOCUS       sequence-317790-        4713 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 4713)
  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 4713)
  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: pNK108h
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      complement(1206..1237)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1253..1912)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(2027..2977)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3025..3247
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(3809..4687)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac ccaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttacctgc
      301 aggtgcagtt ttattttggg agttacacct tggctaacat taagtctgct aagaagcgtg
      361 caatccaagc cgaaaagcgt cgtcagcaca acgctagccg tcgttcaatg actcgtacaa
      421 gcatcaagaa agttatcgct gcaattgcaa gcggtgacaa agaagcggca caagctgcac
      481 ttactgctgc tactccagta cttgacagaa tggctactaa aggcttgatt cacaagaaca
      541 aagctgctcg tcataagagc cgtctagctg cacaaatcaa agcactaggt taatctttac
      601 gacaacgagt ttgaggcatt tatggttact attcgtttac agcgtggtgg cgcgaaaaag
      661 cgtccatttt atcaactagt ggtggctgat agccgtcgtg caagaaacgg tcgtttcatt
      721 gaaaacatcg gtttcttcaa cccaactgca caaggtcaag cggagcgtct tcgcgtagat
      781 ctagagcgtg ttgaatactg ggttggtgta ggcgcgagct tatcagatcg cgtttctagc
      841 ttggtaaaag aagcgaaaaa atctgcagcg taagctgtta tattaatatt aggagaatga
      901 tatgtcacta actaaagcag aaaccgcaaa aatcattgct gagtacggcg ttaaagaagg
      961 cgacactggt tcaccagaag ttcaagttgc tttgcttact cacaacatca acaagcttca
     1021 aggtcacttt gctgatcaca agaaagatca ccactcacgt cgtggtcttc tacgtatggt
     1081 tagccaacgt cgtaagcttc tagactacct taaaggcaag aacgcagagc gttaccttga
     1141 ccttatcaag cgtctaggtc tacgtcgata agactaggta cttaattaac ggcactcctc
     1201 agccaagtca aaagcctccg gtcggaggct tttgactaca tgcccatggc gtttacgccc
     1261 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
     1321 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     1381 tataatattt gcccatagtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
     1441 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
     1501 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
     1561 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
     1621 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
     1681 tcattgccat acggaactcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
     1741 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
     1801 gaacggtctg gttataggta cattgagtaa ctgactgaaa tgcctcaaaa tgttctttac
     1861 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
     1921 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     1981 tatggtgaaa gttggaacct cttacgtgcc aggccaaata ggccgttcag atccttccgt
     2041 atttagccag tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac
     2101 cattgagatc atgcttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct
     2161 gaatttttgc agttaaagca tcgtgtagtg tttttcttag tccgttacgt aggtaggaat
     2221 ctgatgtaat ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt
     2281 cggttacgag atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc
     2341 ggcctcgctt atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg
     2401 gtttcaaaac ccattggtta agccttttaa actcatggta gttattttca agcattaaca
     2461 tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta
     2521 gttcttttaa taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat
     2581 gttccagatt atattttatg aattttttta actggaaaag ataaggcaat atctcttcac
     2641 taaaaactaa ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct
     2701 taaagccttt aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg
     2761 ctttagctaa tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt
     2821 tataagtgaa cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta
     2881 gtgccacaca gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg
     2941 ctagttcatt tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat
     3001 tttaatcact ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt
     3061 gagttgtggg tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta
     3121 gaccctctgt aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta
     3181 taatttatag aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt
     3241 ataactcact actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg
     3301 ctcctctaca aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg
     3361 gcaaatcgct gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt
     3421 cgtgacattc agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac
     3481 aggcgccttt tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg
     3541 ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt
     3601 cagcagttcc tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc
     3661 attcagactg gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt
     3721 actgtcaaga ggacatccgg tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3781 cgttttgctg aggagactta gggacccttt atgacaactt gacggctaca tcattcactt
     3841 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     3901 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     3961 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     4021 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     4081 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     4141 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     4201 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     4261 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     4321 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     4381 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     4441 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     4501 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     4561 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     4621 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     4681 cagccatact tttcatactc ccaccattca gag
//
