LOCUS       sequence-317792-        6716 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 6716)
  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 6716)
  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: pNK131d
FEATURES             Location/Qualifiers
     source          1..6716
                     /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(3209..3240)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3256..3915)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(4030..4980)
                     /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      5028..5250
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(5812..6690)
                     /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 ccgtttgaag gtccgaccaa tggccaataa tcccggcgcc aagaaggcga
      361 tccgcaagat cgcccgccgc accgaagtca acaccgctcg ccgctcgcgc gtgcgcacct
      421 tcctgcgcaa gttcgaagac gcgatcgcca agggcgatgt cgccgtcgcc aaggcggctt
      481 tcgtggaagc tcaatccgag ctgatgcgcg ctgtctccaa gggcgtggtt caccccaaca
      541 cgggctcgcg caaggtctcg cgcttggccg cccgtctgaa gaagctggac aaggccgcag
      601 cctagtcaag attagacctt aaggactacc gcaatgctga agatccgtct cgcccgtggc
      661 ggcgccaaga agcgtccgta ctactcgatc gtcgtcgccg acagccactc gccgcgcgat
      721 ggccgcttca tcgagaaggt cggcacctac aacccgctcc tcaagaagga cgacgccaac
      781 cgcgtcaccc tgaaggtcga gtcgatccag gagtggctca agaagggcgc ccagccgacc
      841 gaccgcgtcg cccgcttcct ggccgcccaa ggcctggtcg cctggacgca cggcaacaac
      901 ccggaaaagg gtaagccggg caagaaggcg caagagcgtc tggccgagcg cgcgcagcgt
      961 gaagaagagc gcaagcaagc cgaagctgac gccaaggccg ccgccgaagc tgaaaaggcc
     1021 gccgctgctg aagccgccgc cgccgccgcc gcggctccgg ccgttgaaga agccccggct
     1081 gaagaggctc cggccgctga agctccggct gaagaagccg ccgaaggcta atcagggttc
     1141 ctagagacca aagatcagca tggctgacga tatgagcttc aacccgacgc gcgacgattt
     1201 cgaagcgctc ctcaacgact ccatgggcgg ccgtgacttc gccgaaggca ccgtcgtcaa
     1261 gggcaaggtc gtcgggatcg aaaaggactt cgccatcatc gacgtcggtc tgaagaccga
     1321 aggccgcgtc cagctgaagg aattcggcgt cgacgagaac ggcaaggcca ccgtgaaggc
     1381 cggcgacacc gtcgaagtct tcctggaacg cctcgaaaac gccatgggcg aagcggtcat
     1441 cagccgcgaa aaggccaagc gcgaagaagc ttggacccgt ctggagggcg tctacgcccg
     1501 caacgagccc gtcatgggcg cgatcgtcgg ccgcgtgaag ggtggcttca ccgtcgacct
     1561 gggcggcgcc tcggccttcc tgccgggcag ccaagtcgac atccgtccgg tccgcgacgt
     1621 cggtccgctg atgggcaagg aacagccctt cgccatcctg aagatggacc gtccgcgcgg
     1681 caacatcgtc gtctcgcgtc gcgccatcct ggaagaagcc cgcgccgagc agcgcaccga
     1741 actggtgtcg cagctgcaag agggtgaaat ccgcgaaggc gtcgtcaaga acatcaccga
     1801 ctacggtgcg ttcgttgacc tgggcggcat cgacggcctg ctgcacgtca ccgacatgag
     1861 ctggaagcgc gtcaaccacc cgagccaagt gctcgccgtg ggcgacaccg tgaaggtcca
     1921 gatcgtcaag atcaacccgg acacccagcg catcagcctc ggcatgaagc agctgcagtc
     1981 ggatccgtgg gacggcgtcg aagccaagta cccggtcggc gccaagttca cgggccgcat
     2041 caccaacatc accgactacg gcgccttcgt ggagctggaa gccggcgttg aaggcctggt
     2101 gcacgtctcg gaaatgtcct ggaccaagaa gaacgttcac cccggcaaga tcgtctcgac
     2161 ctcgcaggaa gtcgacgtcg tggtcctgga cgtcgatccg tccaagcgcc gcgtctcgct
     2221 gggcctgaag caagccctgg ccaacccgtg ggaagccttc ctggaagctc acccggtcgg
     2281 ttcgaccgtc gaaggcgaag tcaagaacgc gaccgagttt ggtctgttcg tcggcctcga
     2341 caacgacatc gacggcatgg tgcacttgtc ggacatcgac tggagcgtcc cgggcgaaga
     2401 agccatggcc cgctacaaga agggcgacat ggtcaaggcg aaggttctcg acgtcgacgt
     2461 cgagaaggaa cgcatctcgc tgggcatcaa gcagctggcc ggcgacccga tgtcgggcga
     2521 cacctaccgc aagaaccaga ccgtgaccgt caccgtcacg gaagtcacct cgggcggcat
     2581 cgaagttcgc ttcggcgaag acgacgctcc gatgaccgcc ttcatccgca agtcggacct
     2641 gtcgcgcgat cgtcaagagc agcgtccgga gcgcttcgcc gtgggtgacc gcgtcgacgc
     2701 tcagatcacc aacgtggaca aggccgcgcg ccgcgtctcg ctgtcgatca agtcgctgga
     2761 aatggccgaa gagaaggaag ccatcgagca gttcggctcg tcggactccg gcgcttcgct
     2821 gggcgacatc ctgggcgcgg ccctgcgcga gcgggcctcc aaggactaat atgaaagacg
     2881 aagcacctga gcggagacaa cccgatgtcg atcactctcg agcgcaaggc cgctcttatc
     2941 gccgaacacg cccgtagcac gggtgacacc ggcagcgccg aagtccaggt cgcgatcctc
     3001 tccgagcgta tcgccaacct gaccgagcac ttcaagacgc acaagaagga caaccacagc
     3061 cgtcgtggcc tgctgaagct ggtttcccag cgtcgccggc ttctcgacca cctgaagaag
     3121 tccgacgccg ctcgctacca gtccctgatc gaaaagctgg gcctgcgtcg ctaaggcatc
     3181 aaacttaatt aacggcactc ctcagccaag tcaaaagcct ccggtcggag gcttttgact
     3241 acatgcccat ggcgtttacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     3301 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     3361 catcagcacc ttgtcgcctt gcgtataata tttgcccata gtgaaaacgg gggcgaagaa
     3421 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     3481 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     3541 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     3601 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     3661 ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag cattcatcag
     3721 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     3781 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag taactgactg
     3841 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     3901 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     3961 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccaggccaa
     4021 ataggccgtt cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt
     4081 ttgcgtgagc catgagaacg aaccattgag atcatgctta ctttgcatgt cactcaaaaa
     4141 ttttgcctca aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct
     4201 tagtccgtta cgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat
     4261 ttttatctgg ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg
     4321 gaaaatcaac gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta
     4381 agtgtttaaa tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg
     4441 gtagttattt tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc
     4501 cttgtgagtt ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta
     4561 tttgttttca aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa
     4621 aagataaggc aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc
     4681 atagtttgtc cactggaaaa tcttaaagcc tttaaccaaa ggattcctga tttccacagt
     4741 tctcgtcatc agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat
     4801 gttcatcatc tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg
     4861 gttttcaatc gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc
     4921 cgttaagtca tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat
     4981 acatctcaat tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg
     5041 ataattacta gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc
     5101 tggaaaactt gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt
     5161 ttttgtttat attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa
     5221 aagaatagat cccagccctg tgtataactc actactttag tcagttccgc agtattacaa
     5281 aaggatgtcg caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt
     5341 aagtagcacc ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca
     5401 ggcacctgag tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag
     5461 tgaatggggg taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca
     5521 taatacaaga aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg
     5581 gtgctatctg actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac
     5641 ttcggattat cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta
     5701 tcatcaacag gcttacccgt cttactgtca agaggacatc cggtcaaata aaacgaaagg
     5761 ctcagtcgaa agactgggcc tttcgttttg ctgaggagac ttagggaccc tttatgacaa
     5821 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     5881 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     5941 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     6001 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     6061 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     6121 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     6181 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     6241 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     6301 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     6361 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     6421 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     6481 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     6541 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     6601 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     6661 cagcagggga tcattttgcg cttcagccat acttttcata ctcccaccat tcagag
//
