LOCUS       sequence-334354-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant F830N.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   Vieni C, Coudray N, Isom GL, Bhabha G, Ekiert DC
  TITLE     Role of Ring6 in the Function of the E. coli MCE Protein LetB.
  JOURNAL   J Mol Biol. 2022 Jan 22;434(7):167463. doi: 
            10.1016/j.jmb.2022.167463.
  PUBMED    35077766
REFERENCE   2  (bases 1 to 7073)
  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: "J Mol 
            Biol."; date: "2022-01-22"; volume: "434(7):167463. doi"; pages: " 
            10.1016/j.jmb.2022.167463"
COMMENT     Sequence Label: pBEL1872
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..46)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    132..271
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     187..206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(286..303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(457..1045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(537..556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1216..2007,2008..2076))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1839..1858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2077..2181)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2249..2267
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2312..2359)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2355..2373
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             6331..6353
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6384..6403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6385..6403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(6912..7073,1..30)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
       61 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      121 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      181 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
      241 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
      301 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      361 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      421 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      481 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      541 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      601 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      661 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      721 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      781 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      841 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
      901 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      961 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1021 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1081 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1141 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1201 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1261 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1321 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1381 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1441 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1501 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     1561 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1621 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1681 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1741 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1801 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     1861 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1921 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1981 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2041 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2101 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2161 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2221 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     2281 agaattatcc ggatatagtt cctcctttca gcaaaaaacc cctcaagacc cgtttagagg
     2341 ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc
     2401 gggctttgtt agcagccgga tctgctcgag gtgggcatta tttgggaagc gcagttcccc
     2461 attcacgcca ctctttcggt tcactttcct gcaacaggaa gtgtttgcct tcctgggctt
     2521 tcggtgccag tggcgtaccc ggaggcgtgg cgaaggcgat gccgccacgg ataaactggt
     2581 tattggtgcc ggttttcact acgccgcccg tcagaccaaa gtccagactg taacccgatg
     2641 ccaaccagaa gacggaattg ttacgcacca ggtgttgata gcgtttactg atgcgcatcg
     2701 caatcatcac gcgatctgac aatgtcccca gcgtcattcc tgtaaccgta ccgacttcca
     2761 gaccacggaa cagcacaggc gtaccgatgc ctaacgaacc ggcttccggc gcttcaacaa
     2821 taatgcttaa gccatccagg taacgcgaat cagtaatggt ggcctcttgt aattcaaagt
     2881 cgcggcgagg attgccccgg cctggttcga cgttgatata cggctggagg atagtatcaa
     2941 gatgctcaac gccagctgcc gaaatttgcg gtgtgaccac tgagaagcgc gtaccaccgc
     3001 gagcaaaggt ctggacatat tccgggtaaa gtaccgcctt tgcctgtact tcattgcgtg
     3061 cggtaatcag atccagcgtc tggatttgcc cgatgtcaat accgagatag cgaattggca
     3121 tcccgaccgc cagttttccg gcatcgaaag cgtgaagcgt aatctgcccg ccaaccgcac
     3181 gggccgctgt ttcggaagca tacagaatac gcttatcgcc tttacgctga ctggcgctgg
     3241 caccgctgag gttgtcgaag ctaatggctc cctttagagc tctggagagc ggggatgcct
     3301 gtacggtcag accactacca ttcagctgaa ctttcgcccc gccttctgcc cagaacacgc
     3361 tattgctggt cagaaggttg cgatactccg gcttaatatg cagatcgatg tcaaacgcgt
     3421 tagctcgcgg acgcacggta atcacttcac caacttcaaa tttacggtac agcactaccg
     3481 atcctgcctg cacatccggc agcgtctctg cactcaaact cacggttgtg gtgggtaaat
     3541 cgctaaggct gttctccagc gctttttcca aattggcata cagtggatag ctggctttca
     3601 tctcaccttt atcgcccggc agaatacgga tcccgccatt gatccattct gaggcgctgg
     3661 caccgagaaa ctcaacgcca tccagcccca ccttcacatc gacacggctg ttgacgacaa
     3721 atttgctatc gccttttacc agttcgcgat gctgaggctc gatggcgacg gtaaaggtga
     3781 cgcctttgct ggtgagttta cgatcgataa cctggcctac ctgcacgccg tgaagaatga
     3841 gcggctgacc cgcatcaata ccgtaacttt ccggcgcggt cagggtcagc gtcagaacat
     3901 caggttcatg cagcagtgct ttttcgcctg gcacaacaac gaattctttg cgtggctcgc
     3961 catcgccggg caccagctcg aaggttttgc cggtcagcag ggcgctgaga ttagcatcgc
     4021 taagggataa tttcgggttg cgtaattcga tgcgggtatt ttcacgaagc agggtaacga
     4081 cgctgggatc aacggtcatt tcccctgtga ctttaccacc aggatttaaa tccagtttag
     4141 tcagctgtcc gacttccagc ccctgataca ttaacggcgt cgagtcggcg gttaatccgg
     4201 ccccactcgg cagttccagt tttattatta cgccacgctg gctgtgggcc agatcttcat
     4261 acagaccaaa ggtatcttcc gcctcggcag gtttcgactc ttctggtgaa tcgaaggcaa
     4321 tcgcaccgtt aaccagtgcc gccaagcttt ccagtttcac cttcgcgcca ctgatactga
     4381 cgttggcatc aacgccggaa acgttccaga aacggctacc ttttttcacc agatcggtaa
     4441 aacgccgctc gatcaggaca tcaatcacca cgccttgctt gttgggattg atggcatagt
     4501 cgtagacttt tcccaccggg atcttgcgga aatagaccaa tgaaccgctg ttcagcgaac
     4561 cgagatcggg ggcttgcagg tggatcatca gatcgccatt gtccagccga tatttcggtt
     4621 gggtatcgag tgcgacaaag tgatcctgct ctttaccttt cccgggcatc atgccgatat
     4681 agttcccacc gacgagggcg tccagcccgg agacacctgc caacgatgct tttggcgtca
     4741 ccagccagaa ctgagtctct tcgcgcagcg catctttcat atcggacttg atgctgacct
     4801 tgacttcaat cttacgaaga tcgtcgctga ggctgatgtc ctgcactgtt ccgacttcaa
     4861 cgccctgata acgaacaggc gtacggcctg gaacaatacc atccgccgac ataaagtcga
     4921 tggtgacggt attaccccgg tcctgataac tgtcccaaat cagccaactg gcaatcatta
     4981 gcgcgatgaa cggcagcagc cagaaaggtg agatacggcg tttattttta atctgcgctt
     5041 cagtcgtcga agcgggcgtt tcctgactca tgtgcatccc aaagtaagcg gctgtccagc
     5101 cattccacag caagaatagt caaaattacc gctgcgccga aataaaacgc agccggtccc
     5161 atagtaaaag cgaggatctg atcgcgatta atcagcgaca tggttaaaga tatgacaaac
     5221 aggtcgagca tcgaccagcg accaatccag gtcaccatcc gcagtaacag aatgcgtgtg
     5281 cgtaaacctt gctggcattt aaaatgaatg ctcaacagta aagtgaacat gacgatcact
     5341 ttagtaaacg ggaccagaat actggcgata aacacgattc ctgcaaccgc aatgttgcta
     5401 cttgccagcg acataattcc ggaaagaatt gtatcttcct gccgtcctcc gttcagataa
     5461 atgatagaaa taggcaacag gttggcaggt aacaataaaa cgattgacgc taacagcgcc
     5521 gcccagcatt tttgcagact atgacggcga cgcaggcgta gcgggatatg gcaacgcggg
     5581 cagcgaccac gctgatctgg atagccggta aaatggcacc ctaggcagac acgaagtttc
     5641 tcgtccctac gcgtagcggg gcgctgcgga taaaatcgct cccacagttc ttcgacatta
     5701 agatgtgaca acgtcaccgt cgttaaaatc accaacgcca caaaagagaa caagccgaca
     5761 ccagcctgga tatgggcata atcctgtacc tttatagaag caacgccaat gccgaccagg
     5821 tagatatcga gcattaccca ctctttaagt cgctcaagca tcagcagtac cggacgtaaa
     5881 ttcattccca gtcggttacc aaaccataaa taagctatgg aggtcaccag aatgagcggg
     5941 gcaccaataa cgcagaaaaa gaccatcgac cccgttatcg catcgccctg tttggtcatt
     6001 tgccagatgc cttgcataac gttggcgtca atacgaatgc ctaacagcca gatatgcaac
     6061 agcggttcgc cccaggcaaa cggcataagc aacagcatgg tgaaggccat tgccgccagg
     6121 cgcgttagcg accagtcgcg cccgtcacga atttttgcct gacagcgcgg acaataggca
     6181 ctttgatgag aatttatctc gggcaggcta aacagcatgt cacattgcgg gcaacgttgg
     6241 taatcaccac gcggcagttc ctcgccgatt gccctcactg ttatcttttt tgtcggcgta
     6301 atttgtggtg tgttaagagc catatgtata tctccttctt aaagttaaac aaaattattt
     6361 ctagagggaa accgttgtgg tctccctata gtgagtcgta ttaatttcgc gggatcgaga
     6421 tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga
     6481 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
     6541 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
     6601 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
     6661 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     6721 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     6781 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     6841 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     6901 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     6961 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     7021 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcg
//
