LOCUS       sequence-317789-        4427 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 4427)
  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 4427)
  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: pNK083d
FEATURES             Location/Qualifiers
     source          1..4427
                     /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(920..951)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(967..1626)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(1741..2691)
                     /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      2739..2961
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(3523..4401)
                     /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 tttttcgagg agctcgacgg tggccaacac accttccgcc aaaaaacgcg
      361 ccaaacaggc tgagaagcgt cgcagccaca acgccagcct gcgctccatg gtgcgcacct
      421 acatcaagaa cgtcgtgaaa gccatcgacg ccaaggacct ggaaaaagcc caggccgcct
      481 tcaccgccgc tgtaccggtg atcgaccgca tggctgacaa aggcatcatc cacaagaaca
      541 aggctgctcg tcataagagc cgtctgagcg gccacatcaa ggccctcagc accgctgccg
      601 cctaacgcac tccactacag gaacgaagtt cacatggtaa ccatccgtct tgctcgtggc
      661 ggctccaaaa agcgcccctt ctaccacctg accgtgacca acagccgcaa tgcgcgcgat
      721 ggtcgcttcg tcgagcgcat cggtttcttc aacccggttg cgactggtgg cgaagtgcgt
      781 ctgtccgtcg accaggagcg tgctacctac tggctcggcc agggtgcaca gccgtctgag
      841 cgcgttgctc agctgctcaa ggacgctgcc aaggcaaacg cctaagccgt catacttaat
      901 taacggcact cctcagccaa gtcaaaagcc tccggtcgga ggcttttgac tacatgccca
      961 tggcgtttac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     1021 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1081 cttgtcgcct tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat
     1141 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1201 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1261 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1321 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1381 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag
     1441 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1501 cgtaatatcc agctgaacgg tctggttata ggtacattga gtaactgact gaaatgcctc
     1561 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1621 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     1681 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccaggcca aataggccgt
     1741 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
     1801 ccatgagaac gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc
     1861 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
     1921 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
     1981 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
     2041 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
     2101 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
     2161 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
     2221 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
     2281 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
     2341 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     2401 ccactggaaa atcttaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     2461 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     2521 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     2581 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     2641 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     2701 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     2761 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     2821 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     2881 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     2941 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     3001 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     3061 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     3121 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     3181 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     3241 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     3301 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     3361 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     3421 ggcttacccg tcttactgtc aagaggacat ccggtcaaat aaaacgaaag gctcagtcga
     3481 aagactgggc ctttcgtttt gctgaggaga cttagggacc ctttatgaca acttgacggc
     3541 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
     3601 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
     3661 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
     3721 gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg
     3781 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
     3841 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
     3901 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
     3961 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
     4021 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
     4081 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
     4141 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
     4201 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
     4261 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
     4321 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
     4381 atcattttgc gcttcagcca tacttttcat actcccacca ttcagag
//
