LOCUS       sequence-317766-        6553 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 6553)
  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 6553)
  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: pAB184d3
FEATURES             Location/Qualifiers
     source          1..6553
                     /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"
     CDS             1262..1273
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(3046..3077)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3093..3752)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             complement(3867..4817)
                     /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      4865..5087
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(5649..6527)
                     /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 aggtgcagta aaatataagg gactcggtga tggctaatca ttcttcagca aaaaaagctg
      361 ttagacaaac agtaaaaaga actctaatca ataaaaaacg atctagtgca ataaagactt
      421 ttataaagaa agttgtacat gaaatcagtc ttggtaataa agaaaatgct aacatagctt
      481 tatctgttgc tcagtctaag ataatgcaag gtgtgaaaaa aaatataatt aaattaaata
      541 ccgcctcaag aaaaatcagc agattatcaa ggcaaataaa aagtttgaaa gtaaataaca
      601 ctttatagta atttaaataa ttgtaagtaa tataatatgg cagtaaaaat tcgtttagct
      661 agaggcggtg ctaaaaagcg tcctttttac cgtgtagtag tagctaacgc aacagcacct
      721 cgtgacggtg actttttaga aaaagtcgga acttatgatc caatgcttgc ttcagataat
      781 agtgagcgtg ttgttttaaa aaaagatcgt ataaaatatt ggcttggcac cggtgctaaa
      841 ccaactgagc gagtcgcaaa atttattgag caagcgggtg ttactcttcc tgagaaagtt
      901 aaaaaggaaa tggaagtgaa agcaaaaaac cgcaaagcta gactaagtaa aaaggaagct
      961 aaagaagctt agtttatagt gctaatacta cgagaagaat atgtcaataa aactaaaaca
     1021 acgatttgtt ccacaacttg cagcaatgaa tcacgaattt gcggaagatt tctctaaaat
     1081 gcttgaaacc gttgatacaa gtcatataaa agaaaaaacg gtagttaaag gtcaagtaat
     1141 cgaaataaaa aacgatataa ttattgttga cgttggatta aaaaatgaag gtagaatacc
     1201 taagtctgaa tttttagctt tacctgaagt tggggacgta gtcgaagttt tcatcgagaa
     1261 aattgaagga cgtaacggca gaacaatatt gagccgtgaa aaagcagtta aagaagaatt
     1321 atggggacaa ctagaaatca tgtgttccaa gggcgaattt gtcgacggta cgattttcgg
     1381 tcgtgtaaaa ggaggcttta ccgtggattt atccggtgta gtagcattct taccgggaag
     1441 ccaagttgac gttagaccta ttaaagatcc tacttctata atgaatatca agcaaccgtt
     1501 taaaatttta agtatggata aaaagcttgg taatatagta gtttcaagaa gagctatttt
     1561 agaagaatca cgttccgaag ctagagatga gatgctatct aaaattaaag aaggcatggt
     1621 attagaagga accgtaaaaa atattactga ttacggtgcg tttatcgatc ttggaagtgt
     1681 tgacggttta ttacatttaa ccgatatttc ttggggaaga gttaaccatc cttcagaagt
     1741 gttagacttt aaccaaaagg ttaaagtaat ggttattaag tttgatgaaa aaacaaaaag
     1801 aatatcactc ggtataaaac aacttgattc taacccatgg gaagcaatta aagaagaatt
     1861 ccccgtcggt aaacaaatga ccggtaaagt tacaaatttt gctgattacg gcgtatttat
     1921 agaattaaaa gacgggcttg agggacttgt tcattcaagc gaaattagtt ggttaaaatc
     1981 taatcaaaat cctagaaaaa cgctaactat agggcaagaa gtagacttcg tagttttaga
     2041 ggtcgatact gaaaagcatc gtgtttcttt aagtattaaa caatgccaag aaaatccttt
     2101 aacaaagttc gctgaaaata atcctgtcgg tactataatt aaagcaccta ttagaaatat
     2161 tacggatttc ggtatttttg ttgcactcgg taataatatg gacggcatga ttcatgaagg
     2221 tgatattagc tgggaagata aaggaacgga tctattaaaa tcatacaaaa aaggtgacga
     2281 aatagaatgc aaagttctag ccattaatat tgaaaaagaa caagttagtt taggtataaa
     2341 acaattgtcg ccgaatccat atcaagaaat cagtgatgaa tataaaaaag gcaccatagt
     2401 taaagccctt ataacagaag ttaaagatga gggactcgaa gtattattaa acgataaagt
     2461 agcaggtttt attaaaagaa ctgaactatc ggatgaaaaa gacgagcaga agcctgaaat
     2521 gtttcaaata gacaaagaaa tagaagccaa agttgtttct atcgagaaat caaccggtag
     2581 aattttatta tcggtaaaag ctcataaaat agcagaacgt caaaaagctc tcaaagaata
     2641 tggttctagt gataatacta ccaatatggg agatatactt gctaatgctt tagaagataa
     2701 gaagtaataa tctattattt aaggagaaat tatcgatgtc gattactaca gaacgtaaac
     2761 aacaattaat taaagaatat gctataacgg aaaatgatac cggttcaagt gcagtgcaat
     2821 gtgctatctt aactgaaaga atcaataatt taaccgagca ttttaaatct aaccataaag
     2881 atcatacttc aagacgcgga ttattaattt tagttgggcg ccgccgcaga ttacttaact
     2941 atattaaaaa gaataatgtt agcaaatatt tagatttaat aagtaagcta ggaattagaa
     3001 agattaagta gttggataaa cttaattaac ggcactcctc agccaagtca aaagcctccg
     3061 gtcggaggct tttgactaca tgcccatggc gtttacgccc cgccctgcca ctcatcgcag
     3121 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     3181 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg
     3241 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     3301 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     3361 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     3421 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     3481 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc
     3541 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     3601 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     3661 cattgagtaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     3721 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     3781 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     3841 cttacgtgcc aggccaaata ggccgttcag atccttccgt atttagccag tatgttctct
     3901 agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt
     3961 tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca
     4021 tcgtgtagtg tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt
     4081 attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt
     4141 ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc
     4201 aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta
     4261 agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg
     4321 ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca
     4381 taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg
     4441 aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt
     4501 tcgcttgaga acttggcata gtttgtccac tggaaaatct taaagccttt aaccaaagga
     4561 ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa
     4621 gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc
     4681 cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt
     4741 agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa
     4801 acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg
     4861 agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa
     4921 ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct
     4981 agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa
     5041 gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact actttagtca
     5101 gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct
     5161 taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct
     5221 tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc
     5281 gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca
     5341 tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat
     5401 ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga
     5461 ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac
     5521 ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcaaga ggacatccgg
     5581 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttgctg aggagactta
     5641 gggacccttt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     5701 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     5761 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     5821 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     5881 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     5941 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     6001 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     6061 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     6121 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     6181 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     6241 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     6301 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     6361 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     6421 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     6481 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     6541 ccaccattca gag
//
