LOCUS       sequence-330959-        6530 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 6530)
  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 6530)
  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: pNK132d
FEATURES             Location/Qualifiers
     source          1..6530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        81..365
                     /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     288..307
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      complement(3103..3134)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3150..3809)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(3924..4874)
                     /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      4922..5144
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(join(5706..6530,1..54))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
       61 catactccca ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca
      121 ctgcgtcttt tactggctct tctcgctaac ccaaccggta accccgctta ttaaaagcat
      181 tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca
      241 cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt
      301 tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc
      361 tccatacccg ttttacctgc aggtgcagtg ctgcaggatt tgagagatca tggctaatac
      421 gtcttccgcc aagaaagcga cccgcaagat cgcccggcgc accgcggtga acaagtcgcg
      481 tcggactcag atgcgcgggt cggtccggat cgtcgaagag gcgatcgcca gcggcgaccg
      541 cgatgcggcg ctgaaggcga tggctcgcgc cgagccggag ctgatgcgcg ccgctcagcg
      601 caacatcatc caccggaacg ccgcgagccg gaaggtgtcg cgtctgacgc acagcatcgc
      661 caagctggcg aagtaattct caccacccca cctaaggagc taacatgtcc gtcgttatcc
      721 gcctcgcgcg cgctggtacc aagaagcgcc cgttctatca tgtcgtcgtc gccgactcgc
      781 gcttcccgcg cgatggccgc ttcatcgagc gtctcggcta cttcaacccg ctgatggcga
      841 aggacaacga agcccgcctg aagctcgacc tcgacaaggt caaggattgg ctcgccaagg
      901 gcgcgcagcc gtcggatcgc gtcgcccgct tcctcgacac cgccggtgtc cgcaagcgtg
      961 aagcccgcaa caaccccgag aaggcggtgc cgcgcaagga gcgcaaggcc gccgacggca
     1021 agtaagcggg gcgattgatg ccgtccatcc gcatctggag aacaaatggc ttcgactgac
     1081 acttacaacc cgacccgcga cgatttcgcc gcgatgctcg acgagtcctt cgccggcggc
     1141 aacctgcagg aaagctctgt tatcaagggc aaggtcgtcg cgatcgagaa ggacatggcc
     1201 gtcattgacg tcggcctgaa gaccgaaggc cgcgtgccgc tgcgcgaatt cgcaggtccg
     1261 ggccgcgaca acgagatcaa ggtcggcgat accgtcgagg tgttcctcga tcggatcgag
     1321 aatgcgctcg gcgaagccgt gctgtcgcgc gacaaggcgc gccgcgaaga gagctggggc
     1381 aagctcgaga aggccttcca gaacaacgag aaggtgttcg gcgtcatctt caatcaggtc
     1441 aagggcggct tcacggtcga cctcgacggc gccgtggcgt tcctgccgcg ctcgcaggta
     1501 gacattcgtc cgatccgcga cgttgcgccg ctgatgaaca actcgcagcc gttccagatc
     1561 ctcaagatgg atcgccgccg tggcaacatc gtcgtgtcgc gccggaccgt gctggaagag
     1621 acccgcgccg agcagcgcca ggagctggtg cagaacctcg aagagggtca ggtgatcgac
     1681 ggcgtcgtca agaacatcac cgattacggt gcgttcgtcg acctcggcgg catcgacggc
     1741 ctgctgcacg tgaccgatat cgcctggcgc cgggtcaatc acccgaccga ggtgctgacc
     1801 atcggccaga ccgtcaaggt caagatcatc aagatcaacc acgagaccca ccgcatctcg
     1861 ctcggcatga agcagctgct ggacgatccg tggcagggca tcgaggccaa gtacccgctg
     1921 aacgcgcgct tcaccggccg cgtgaccaac atcaccgact acggtgcgtt cgtcgagctc
     1981 gagccgggca tcgaaggcct gatccacgtc tccgagatgt cgtggaccaa gaagaacatg
     2041 caccccggca agatcgtttc gacctctcag gaagtcgaag tgcaggtgct cgaggtggat
     2101 tcggtcaagc gccgtatctc gctcggtctc aagcagacca tgcgcaatcc gtgggaagtg
     2161 ttcgtcgaga agcatccggt cggttcgacc gtcgaaggcg aggtcaagaa caagaccgag
     2221 ttcggtctgt tcctgggtct cgacggcgac gtcgacggca tggtccacct ctcggacctc
     2281 gactggaagc tgccgggcga gcaggtgatc gacaacttca agaagggcga catggtcaag
     2341 gccgtggtgc tcgacgtgga cgtcgaaaag gagcgcatct cgctcggcgt caagcagctc
     2401 gaaggcgacc ccttcgccga gccgggcgac gtcaagaagg gtgctgtcgt gacctgcgaa
     2461 gtgctcgacg tgaaggaaag cggcatcgac gtgcagatcg tcggcaccga cttcaacacc
     2521 ttcatcaagc gctcggagct ggcgcgcgac cgcaacgacc agcgcagcga ccgcttcgcg
     2581 gttggtgaga aggtcgacgc ccgcgtcatc cagttcgaca agaaggcccg caaggtccag
     2641 gtgtcgatca aggcgctgga agtggccgag gagaaggaag ccatcgcgca gtacggctcg
     2701 tccgattcgg gcgcgacgct gggcgacatt ctcggcaccg cgctcaagca gcgcgacaag
     2761 taagcctcag gcgcaaggaa agtatctgac gatgtcgatt accgcagaac gcaaagcgga
     2821 agtcatcaag accagcgcca ccaaggccgg tgataccggt tcgcccgagg ttcaggtcgc
     2881 gatcctgtcg gaacgcatca ccaacctgac cgcgcacttc aagacccaca ccaaggacaa
     2941 ccactcccgc cgtgggctgt tgaagctggt gtcgacgcgc cgttcgcttc tcgactacat
     3001 caagaagaag gacgaagcgc gctacaaggc gctgctcgag aagcacaaca tccgtcgctg
     3061 aggcgtcgga cgaccgactt aattaacggc actcctcagc caagtcaaaa gcctccggtc
     3121 ggaggctttt gactacatgc ccatggcgtt tacgccccgc cctgccactc atcgcagtac
     3181 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     3241 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catagtgaaa
     3301 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     3361 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     3421 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     3481 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     3541 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg gaactccgga
     3601 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     3661 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     3721 tgagtaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     3781 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     3841 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     3901 acgtgccagg ccaaataggc cgttcagatc cttccgtatt tagccagtat gttctctagt
     3961 gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcatg cttactttgc
     4021 atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt taaagcatcg
     4081 tgtagtgttt ttcttagtcc gttacgtagg taggaatctg atgtaatggt tgttggtatt
     4141 ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc catttgtcta
     4201 tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc aaccaccaat
     4261 ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca ttggttaagc
     4321 cttttaaact catggtagtt attttcaagc attaacatga acttaaattc atcaaggcta
     4381 atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa ccactcataa
     4441 atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata ttttatgaat
     4501 ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc taatttttcg
     4561 cttgagaact tggcatagtt tgtccactgg aaaatcttaa agcctttaac caaaggattc
     4621 ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac accataagca
     4681 ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga taccgtccgt
     4741 tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca taaaattagc
     4801 ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc tttgaaaaca
     4861 actaattcag acatacatct caattggtct aggtgatttt aatcactata ccaattgaga
     4921 tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat ctgtaaattc
     4981 tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa ttccgctaga
     5041 cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat aaagaaagaa
     5101 taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact ttagtcagtt
     5161 ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa cagaccttaa
     5221 aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa tattcctttt
     5281 gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt tcgctgcgct
     5341 cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat ggattcatgc
     5401 aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc gttttatggc
     5461 gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt
     5521 tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca
     5581 gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtcaagagga catccggtca
     5641 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttgctgagg agacttaggg
     5701 accctttatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
     5761 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
     5821 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
     5881 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
     5941 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
     6001 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
     6061 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
     6121 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
     6181 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
     6241 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
     6301 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
     6361 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
     6421 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
     6481 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
//
