LOCUS       sequence-317705-        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses heterologous rRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  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 7418)
  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: pRF004d
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(105..122)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      274..360
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      452..479
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        539..643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(644..712,713..1504)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPASWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(862..881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1674..2262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2163..2182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2349..2404
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    2355..2373
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2385..2403
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(4058..4072)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="MNHKV"
ORIGIN
        1 ggtgtgtaag tgtagcgata cattgagcta accgatacta atgaaccgtg aggcttaacc
       61 ttacaacgcc gaagctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
      121 tcagaacgca gaagcggtct gataaaacag aattgcctgg cggctgtagc gcggtggtcc
      181 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
      241 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
      301 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      361 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
      421 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
      481 gcgtttctac aaactcttcc tgtcgtcata tctacaagcc ggcgcgccgg gaaatgtgcg
      541 cggaaccctt atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      601 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      661 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      721 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      781 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      841 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
      901 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      961 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgcaataac
     1021 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1081 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1141 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     1201 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1261 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctag
     1321 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1381 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1441 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1501 gtaactgcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1801 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     1861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2281 tttttacggt tcctggcctt ttgctggaga cttagggacc ctgcggccgc cattaattcc
     2341 taatttttgt tgacactcta tcattgatag agttatttta ccactcccta tcagtgatag
     2401 agaaaagtga aatgaataga ccggccgctg agaaaaagcg aagcggcact gctctttaac
     2461 aatttatcag acaatctgtg tgggcactcg aagatacgga ttcttaacgt cgcaagacga
     2521 aaaatgaata ccaagtctca agagtgaaca cgtaattcat tacgaagttt aattctttga
     2581 gcgtcaaact tttaaattga agagtttgat catggctcag attgaacgct ggcggcaggc
     2641 ctaacacatg caagtcgaac ggtagcacag agagcttgct ctcgggtgac gagtggcgga
     2701 cgggtgagta atgtctggga aactgcccga tggaggggga taactactgg aaacggtagc
     2761 taataccgca taacgtcgca agaccaaaga gggggacctt cgggcctctt gccatcggat
     2821 gtgcccagat gggattagct agtaggtggg gtaacggctc acctaggcga cgatccctag
     2881 ctggtctgag aggatgacca gccacactgg aactgagaca cggtccagac tcctacggga
     2941 ggcagcagtg gggaatattg cacaatgggc gcaagcctga tgcagccatg ccgcgtgtat
     3001 gaagaaggcc ttcgggttgt aaagtacttt cagcgaggag gaaggcgttg tggttaataa
     3061 ccgcaacgat tgacgttact cgcagaagaa gcaccggcta actccgtgcc agcagccgcg
     3121 gtaatacgga gggtgcaagc gttaatcgga attactgggc gtaaagcgca cgcaggcggt
     3181 ctgtcaagtc ggatgtgaaa tccccgggct caacctggga actgcatccg aaactggcag
     3241 gctagagtct tgtagagggg ggtagaattc caggtgtagc ggtgaaatgc gtagagatct
     3301 ggaggaatac cggtggcgaa ggcggccccc tggacaaaga ctgacgctca ggtgcgaaag
     3361 cgtggggagc aaacaggatt agataccctg gtagtccacg ccgtaaacga tgtcgacttg
     3421 gaggttgtgc ccttgaggcg tggcttccgg agctaacgcg ttaagtcgac cgcctgggga
     3481 gtacggccgc aaggttaaaa ctcaaatgaa ttgacggggg cccgcacaag cggtggagca
     3541 tgtggtttaa ttcgatgcaa cgcgaagaac cttacctact cttgacatcc agagaactta
     3601 gcagagatgc tttggtgcct tcgggaactc tgagacaggt gctgcatggc tgtcgtcagc
     3661 tcgtgttgtg aaatgttggg ttaagtcccg caacgagcgc aacccttatc ctttgttgcc
     3721 agcggtccgg ccgggaactc aaaggagact gccagtgata aactggagga aggtggggat
     3781 gacgtcaagt catcatggcc cttacgagta gggctacaca cgtgctacaa tggcatatac
     3841 aaagagaagc gacctcgcga gagcaagcgg acctcataaa gtatgtcgta gtccggattg
     3901 gagtctgcaa ctcgactcca tgaagtcgga atcgctagta atcgtggatc agaatgccac
     3961 ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac accatgggag tgggttgcaa
     4021 aagaagtagg tagcttaacc ttcgggaggg cgcttaccac tttgtgattc atgactgggg
     4081 tgaagtcgta acaaggtaac cgtaggggaa cctgcggttg gatcatgtgg ttaccttaaa
     4141 gaagcgtact ttgtagtgct cacacagatt gtctgataga aagtgaaaag caaggcgttt
     4201 acgcgttggg agtgaggctg aagagaataa ggccgttcgc tttctattaa tgaaagctca
     4261 ccctacacga aaatatcacg caacgcgtga taagcaattt tcgtgtcccc ttcgtctaga
     4321 ggcccaggac accgcccttt cacggcggta acaggggttc gaatccccta ggggacgcca
     4381 cttgctggtt tgtgagtgaa agtcgccgac cttaatatct caaaactcat cttcgggtga
     4441 tgtttgagat atttgctctt taaaaatctg gatcaagctg aaaattgaaa cactgaacaa
     4501 cgagagttgt tcgtgagtct ctcaaatttt cgcaacacga tgatgaatcg aaagaaacat
     4561 cttcgggttg tgaggttaag cgactaagcg tacacggtgg atgccctggc agtcagaggc
     4621 gatgaaggac gtgctaatct gcgataagcg tcggtaaggt gatatgaacc gttataaccg
     4681 gcgatttccg aatggggaaa cccagtgtga ttcgtcacac tatcgttaac tgaatacata
     4741 ggttaacgaa gcgaaccggg ggaactgaaa catctaagta ccccgaggaa aagaaatcaa
     4801 ccgagattcc cccagtagcg gcgagcgaac ggggagcagc ccagagtctg aatcagcttg
     4861 tgtgttagtg gaacggtctg gaaagtccga cggtacaggg tgatagtccc gtacaccaaa
     4921 atgcacaggt tgtgaactcg aagagtaggg cgggacacgt ggtatcctgt ctgaatatgg
     4981 ggggaccatc ctccaaggct aaatactcct gactgaccga tagtgaacca gtaccgtgag
     5041 ggaaaggcga aaagaacccc ggcgagggga gtgaaaaaga acctgaaacc gtgtacgtac
     5101 aagcagtggg agcctcttta tggggtgact gcgtaccttt tgtataatgg gtcagcgact
     5161 tatattctgt agcaaggtta accgtatagg ggagccgaag ggaaaccgag tcttaactgg
     5221 gcgttaagtt gcagggtata gacccgaaac ccggtgatct agccatgggc aggttgaagg
     5281 ttgggtaaca ctaactggag gaccgaaccg actaatgttg aaaaattagc ggatgacttg
     5341 tggctggggg tgaaaggcca atcaaaccgg gagatagctg gttctccccg aaagctattt
     5401 aggtagcgcc tcgtgaactc atcttcgggg gtagagcact gtttcggcta gggggtcatc
     5461 ccgacttacc aacccgatgc aaactgcgaa taccgaagaa tgttatcacg ggagacacac
     5521 ggcgggtgct aacgtccgtc gtgaagaggg aaacaaccca gaccgccagc taaggtccca
     5581 aagtcatggt taagtgggaa acgatgtggg aaggcacaga cagccaggat gttggcttag
     5641 aagcagccat catttaaaga aagcgtaata gctcactggt cgagtcggcc tgcgcggaag
     5701 atgtaacggg gctaaaccat gcaccgaagc tgcggcagcg acactatgtg ttgttgggta
     5761 ggggagcgtt ctgtaagccg ttgaaggtgt gctgtgaggc atgctggagg tatcagaagt
     5821 gcgaatgctg acataagtaa cgataatgca ggtgaaaaac ccgcacgccg gaagaccaag
     5881 ggttcctgtc caacgttaat cggggcaggg tgagtcgacc cctaaggcga ggccgaaagg
     5941 cgtagtcgat gggaaacagg ttaatattcc tgtacttggt gttactgcga aggggggacg
     6001 gagaaggcta tgttagccgg gcgacggttg tcccggttta agcatgtagg cggaggttcc
     6061 aggtaaatcc ggtacctttt taacgctgag gtgtgatgac gaggcactac ggtgctgaag
     6121 taacaaatgc cctgcttcca ggaaaagcct ctaagcatca ggtaacacga aatcgtaccc
     6181 caaaccgaca caggtggtca ggtagagaat accaaggcgc ttgagagaac tcgggtgaag
     6241 gaactaggca aaatggtgcc gtaacttcgg gagaaggcac gctgatatgt aggtgaagtg
     6301 atttactcat ggagctgaaa tcagtcgaag ataccagctg gctgcaactg tttattaaaa
     6361 acacagcact gtgcaaacac gaaagtggac gtatacggtg tgacgcctgc ccggtgccgg
     6421 aaggttaatt gatggggtta tccgtaagga gaagctcttg atcgaagccc cggtaaacgg
     6481 cggccgtaac tataacggtc ctaaggtagc gaaattcctt gtcgggtaag ttccgacctg
     6541 cacgaatggc gtaatgatgg ccaggctgtc tccacccgag actcagtgaa attgaactcg
     6601 ctgtgaagat gcagtgtacc cgcggcaaga cggaaagacc ccgtgaacct ttactatagc
     6661 ttgacactga acactggtcc ttgatgtgta ggataggtgg gaggctttga agtgtggacg
     6721 ccagtctgca tggagccatc cttgaaatac caccctttaa tggctggtgt tctaacgtgg
     6781 acccgttacc cgggttgcgg acagtgtctg gtgggtagtt tgactggggc ggtctcctcc
     6841 taaagagtaa cggaggagca cgaaggttag ctaatcctgg tcggacatca ggaggttagt
     6901 gcaaaggcat aagctagctt gactgcgaga gtgacggctc gagcaggtgc gaaagcaggt
     6961 cttagtgatc cggtggttct gaatggaagg gccatcgctc aacggataaa aggtactccg
     7021 gggataacag gctgataccg cccaagagtt catatcgacg gcggtgtttg gcacctcgat
     7081 gtcggctcat cacatcctgg ggctgaagta ggtcccaagg gtatggctgt tcgccattta
     7141 aagtggtacg cgagctgggt ttagaacgtc gtgagacagt tcggtcccta tctgccgtgg
     7201 gcgctggaga attgaggggg gctgctccta gtacgagagg accggagtgg acgcatcact
     7261 ggtgttcggg ttgtcatgcc aatggcattg cccggtagct aaatgcggaa gagataagtg
     7321 ctgaaagcat ctaagcacga aacttgcccc gagatgagtt ctccctgaga ctataagtct
     7381 cctgaaggaa cgttgaagac gacgacgttg ataggtcg
//
