LOCUS       sequence-301855-        7231 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 7231)
  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 7231)
  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: pAC040
FEATURES             Location/Qualifiers
     source          1..7231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..97
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    48..66
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    78..96
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3072..3083
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5029..5046)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5198..5284
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5376..5403
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5463..5567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5568..5636,5637..6428)
                     /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(5786..5805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6598..7186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7087..7106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agacttaggg accctgcggc cgccattaat tcctaatttt tgttgacact ctatcattga
       61 tagagttatt ttaccactcc ctatcagtga tagagaaaag tgaaatgaat agaccggccg
      121 ctgagaaaaa gcgaagcggc actgctcttt aacaatttat cagacaatct gtgtgggcac
      181 tcgaagatac ggattcttaa cgtcgcaaga cgaaaaatga ataccaagtc tcaagagtga
      241 acacgtaatt cattacgaag tttaattctt tgagcgtcaa acttttaaac ttgagagttt
      301 gatcctggct cagaacgaac gctatcggta tgcttaacac atgcaagtcg aacggactaa
      361 tttggggctt gctccaatta gttagtggca gacgggtgag taacacgtgg gaatctaccc
      421 attagtacgg aataactttt agaaataaaa gctaataccg tatattctct gcggaggaaa
      481 gatttatcgc tgatggatga gcccgcgtca gattaggtag ttggtgaggt aatagctcac
      541 caagccgacg atctgtagct ggtctgagag gatgatcagc cacactggga ctgagacacg
      601 gcccagactc ctacgggagg cagcagtggg gaatattgga caatgggcga aagcctgatc
      661 cagcaatacc gagtgagtga tgaaggcctt agggttgtaa agctctttta gcaaggaaga
      721 taatgacgtt acttgcagaa aaagccccgg ctaactccgt gccagcagcc gcggtaagac
      781 ggagggggct agcgttgttc ggaattactg ggcgtaaaga gtgcgtaggc ggtttagtaa
      841 gttggaagtg aaagcccggg gcttaacctc ggaattgctt tcaaaactac taatctagag
      901 tgtagtaggg gatgatggaa ttcctagtgt agaggtgaaa ttcttagata ttaggaggaa
      961 caccggtggc gaaggcggtc gtctgggcta caactgacgc tgatgcacga aagcgtgggg
     1021 agcaaacagg attagatacc ctggtagtcc acgccgtaaa cgatgagtgc tagatatcgg
     1081 aagattctct ttcggtttcg cagctaacgc attaagcact ccgcctgggg agtacggtcg
     1141 caagattaaa actcaaagga attgacgggg gctcgcacaa gcggtggagc atgcggttta
     1201 attcgatgtt acgcgaaaaa ccttaccaac ccttgacatg gtggtcgcgg atcgcagaga
     1261 tgcttttctt cagctcggct ggaccacaca caggtgttgc atggctgtcg tcagctcgtg
     1321 tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc tcattcttat ttgccagcgg
     1381 gtaatgccgg gaactataag aaaactgccg gtgataagcc ggaggaaggt ggggacgacg
     1441 tcaagtcatc atggccctta cgggttgggc tacacgcgtg ctacaatggt gtttacagag
     1501 ggaagcaaga cggcgacgtg gagcaaatcc ctaaaagaca tctcagttcg gattgttctc
     1561 tgcaactcga gagcatgaag ttggaatcgc tagtaatcgc ggatcagcat gccgcggtga
     1621 atacgttctc gggccttgta cacactgccc gtcacgccat gggagttagt tttacctgaa
     1681 ggtggtgagc taacgcaaga ggcagccaac cacggtaaaa ttagcgactg gggtgaagtc
     1741 gtaacaaggt agccgtaggg gaacctgcgg ctggattatg tggttacctt aaagaagcgt
     1801 actttgtagt gctcacacag attgtctgat agaaagtgaa aagcaaggcg tttacgcgtt
     1861 gggagtgagg ctgaagagaa taaggccgtt cgctttctat taatgaaagc tcaccctaca
     1921 cgaaaatatc acgcaacgcg tgataagcaa ttttcgtgtc cccttcgtct agaggcccag
     1981 gacaccgccc tttcacggcg gtaacagggg ttcgaatccc ctaggggacg ccacttgctg
     2041 gtttgtgagt gaaagtcgcc gaccttaata tctcaaaact catcttcggg tgatgtttga
     2101 gatatttgct ctttaaaaat ctggatcaag ctgaaaattg aaacactgaa caacgagagt
     2161 tgttcgtgag tctctcaaat tttcgcaaca cgatgatgaa tcgaaagaaa catcttcggg
     2221 ttgtgaagta aatataataa gagcatttgg tggatgcctt ggcactagaa ggcgatgaag
     2281 gacgtaatac gctgcgataa gcttcgggga gttgcgaata aactttgatc cgaagatttc
     2341 cgaatagggg aacctaccta atttattagg tattgtatag tgaatacata gctatatgag
     2401 gcaaacccag cgaactgaaa tatctaagta gctggaggaa aggacatcaa ccgagactcc
     2461 gttagtagtg acgagcgaac gcggatcagg ccagtggctt caaaaaaata actaaaacaa
     2521 catggaaagg ttgaccatag agggtgatag tcccgtatag gtaaaaagtt tgaagtcctt
     2581 gagtaaggcg ggacacgtga aatcctgttt gaacataggg ggaccaccct ctaagcctaa
     2641 gtactctcta gtgaccgata gtgaacaagt accgtgaggg aaaggtgaaa agtaccccga
     2701 caaggggagt gaaatagtat ctgaaaccaa atgcttacaa gcagtcggag caagcgtatt
     2761 tatatgtctt gtgacggcgt accttttgta taatgggtca gcgacttagt ttatctagca
     2821 agcttaagcc gttaggtgta ggcgtagcga aagcgagtct gaatagggcg tttagttaga
     2881 tgaattagac ccgaaaccgg gtgatctagc catggccagg ttgaaagcgg agtaaaatcc
     2941 actgaaggac cgaacccact actgttgaaa aagtagggga tgagctgtgg ttaggggtga
     3001 aaggccaatc aaactcggat atagctggtt ctccgcgaaa tctatttagg tagagcgtta
     3061 tagcgattac catcgaaggt agagcactga atgagctagg gggtcccaca gacttaccaa
     3121 actcaatcaa actccgaatg tcgatgagta cagcatagca gacagactat gggtgctaag
     3181 gtccatagtc gagagggaaa aagcccagac cgccatctaa ggtccctaaa tcatgactaa
     3241 gtgttaaagg atgtgagaag accaaaacaa ctaggatgtt ggcttagaag cagccatcat
     3301 ttaaagaaag cgtaatagct cactagtcta aataagtttt cttgcgccaa caatgtaacg
     3361 gggctcaagt catgtaccga agatgcgggt ttatgcattt ggcataaatg gtagcggagc
     3421 gttccgtaag cctgcgaagg tgaaccgtga ggtttgctgg aggtatcgga agtgagaatg
     3481 ctgacataag tagcgataaa gaatgtgaga aacattctcg ccgaaagtcc aagggttctt
     3541 gcgtaaagtt aatctgcgca aggttagtcg gcccctaaga tgaggctgaa aggcgtaatc
     3601 gatgggaatc aggttaatat tcctgaacct gaaggatgtg acgaaaatag taaattgtac
     3661 agtcttattg gattgattgt gcggtgaata ttttccagga aataacacct ttattataga
     3721 ccgtacccga aaccgacaca ggtggacagg tagagtatac caaggcgctt gagagaacga
     3781 tgctgaagga actaggcaaa ttgcatctgt aacttcggaa gaaagatgac ctgtatatgg
     3841 gcaaccatgt gcaggtggca caagctaggg ggtagcgact gtttattaaa aacacagggc
     3901 tctgcaaagt caatagacga tgtatagggt ctgacgcctg cccagtgctg gaagattaaa
     3961 aggaggggtg caagctctga attgaagtcc cagtgaacgg cggccgtaac tatgacggtc
     4021 ctaaggtagc gaaattcctt gtcgggtaag ttccgacccg cacgaatggc gtaacgattt
     4081 ccccactgtc tccagtatcg actcagcgaa attgaattct ccgtgaagat gcggagttcc
     4141 cgcggtcaga cggaaagacc ccgtgaacct ttactatagc tttgcactgg tgttaggaat
     4201 cagatgtgca ggataggtgg gagactacga agcgaaggcg tcagcctttg tggagtcacc
     4261 cttgagatac cacccttttg gtgcttgaca tctaaccgag atccttgaat caggatccga
     4321 gacaatgcat ggtgggtagt ttgactgggg cggtcgcctc ccaaagagta acggaggcgc
     4381 gcgatggtta gctcaggttg gtcggaaatc aacttttaga gtgcaatggc ataagctagc
     4441 ctgactgcga gtctgacaag acgagcagag acgaaagtcg gtcatagtga tccggtggtc
     4501 ccgagtggaa gggccatcgc tcaacgaata aaaggtactc cggggataac aggctgatga
     4561 tttccaagcg tccatagcga cgaaatcgtt tggcacctcg atgtcggctc atcacatcct
     4621 ggggctggag aaggtcccaa gggttcggct gttcgccgat taaagtggta cgtgagctgg
     4681 gtttagaacg tcgtgagaca gttcggtccc tatctgccgt gggtgtagga agtttgagag
     4741 gatctgcctt tagtacgaga ggaccgaggt ggacgtaccc ctggtggacc agttgtcgtg
     4801 ccaacggcac agctgggtag ctaagtacgg aagggataac tgctgaatgc atctaagcag
     4861 gaaacccacc tcaaaactag acttccccat tagagccgtg gaagaccacc acgttgatag
     4921 gtcggatgtg gaagcacagt aatgtgtgta gctaaccgat actaatagct cgattgattt
     4981 actttgacaa cgccgaagct gttttggcgg atgagagaag attttcagcc tgatacagat
     5041 taaatcagaa cgcagaagcg gtctgataaa acagaattag cttggtggtt atagcatgag
     5101 tgaaacacac gatcccatcc cgaactcgaa tgtgaaacct catagcgcta atggtactat
     5161 gtcataaggc atgggagagt aagtcgctgc caagcttcaa ataaaacgaa aggctcagtc
     5221 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     5281 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     5341 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     5401 ttttgcgttt ctacaaactc ttcctgtcgt catatctaca agccggcgcg ccgggaaatg
     5461 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5521 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5581 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5641 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5701 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5761 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     5821 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5881 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcaa
     5941 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6001 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6061 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     6121 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6181 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6241 ctagctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6301 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6361 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6421 attggtaact gcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6481 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6541 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6601 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6661 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6721 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     6781 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6841 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6901 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6961 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7021 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7081 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7141 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7201 ggccttttta cggttcctgg ccttttgctg g
//
