LOCUS       sequence-345241-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB(A706N).
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: pBEL2417
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     257..274
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    289..428
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(354..373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     514..536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(530..721)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1230..1249
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1230..1248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1280..1302
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5260..5278)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5274..5321
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5366..5384)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5452..5556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5557..5625,5626..6417)
                     /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     complement(5775..5794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6588..7073,1..103)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
       61 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      121 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
      181 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
      241 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
      301 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
      361 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
      421 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      481 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      541 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
      601 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
      661 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
      721 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
      781 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
      841 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
      901 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
      961 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     1021 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     1081 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     1141 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     1201 cgctgcccga gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
     1261 gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata tggctcttaa
     1321 cacaccacaa attacgccga caaaaaagat aacagtgagg gcaatcggcg aggaactgcc
     1381 gcgtggtgat taccaacgtt gcccgcaatg tgacatgctg tttagcctgc ccgagataaa
     1441 ttctcatcaa agtgcctatt gtccgcgctg tcaggcaaaa attcgtgacg ggcgcgactg
     1501 gtcgctaacg cgcctggcgg caatggcctt caccatgctg ttgcttatgc cgtttgcctg
     1561 gggcgaaccg ctgttgcata tctggctgtt aggcattcgt attgacgcca acgttatgca
     1621 aggcatctgg caaatgacca aacagggcga tgcgataacg gggtcgatgg tctttttctg
     1681 cgttattggt gccccgctca ttctggtgac ctccatagct tatttatggt ttggtaaccg
     1741 actgggaatg aatttacgtc cggtactgct gatgcttgag cgacttaaag agtgggtaat
     1801 gctcgatatc tacctggtcg gcattggcgt tgcttctata aaggtacagg attatgccca
     1861 tatccaggct ggtgtcggct tgttctcttt tgtggcgttg gtgattttaa cgacggtgac
     1921 gttgtcacat cttaatgtcg aagaactgtg ggagcgattt tatccgcagc gccccgctac
     1981 gcgtagggac gagaaacttc gtgtctgtct tgggtgccat tttaccggct atccagatca
     2041 gcgtggtcgc tgcccgcgtt gccatatccc gctacgcctg cgtcgccgtc atagtctgca
     2101 aaaatgctgg gcggcgctgt tagcgtcaat cgttttattg ttacctgcca acctgttgcc
     2161 tatttctatc atttatctga acggaggacg gcaggaagat acaattcttt ccggaattat
     2221 gtcgctggca agtagcaaca ttgcggttgc aggaatcgtg tttatcgcca gtattctggt
     2281 accgtttact aaagtgatcg tcatgttcac tttactgttg agcattcatt ttaaatgcca
     2341 gcaaggttta cgcacacgca ttctgttact gcggatggtg acctggattg gtcgctggtc
     2401 gatgctcgac ctgtttgtca tatctttaac catgtcgctg attaatcgcg atcagatcct
     2461 cgcttttact atgggaccgg ctgcgtttta tttcggcgca gcggtaattt tgactattct
     2521 tgctgtggaa tggctggaca gccgcttact ttgggatgca catgagtcag gaaacgcccg
     2581 cttcgacgac tgaagcgcag attaaaaata aacgccgtat ctcacctttc tggctgctgc
     2641 cgttcatcgc gctaatgatt gccagttggc tgatttggga cagttatcag gaccggggta
     2701 ataccgtcac catcgacttt atgtcggcgg atggtattgt tccaggccgt acgcctgttc
     2761 gttatcaggg cgttgaagtc ggaacagtgc aggatatcag cctcagcgac gatcttcgta
     2821 agattgaagt caaggtcagc atcaagtccg atatgaaaga tgcgctgcgc gaagagactc
     2881 agttctggct ggtgacgcca aaagcatcgt tggcaggtgt ctccgggctg gacgccctcg
     2941 tcggtgggaa ctatatcggc atgatgccgg gtaaaggtaa agagcaggat cactttgtcg
     3001 cactcgatac ccaaccgaaa tatcggctgg acaatggcga tctgatgatc cacctgcaag
     3061 cccccgatct cggttcgctg aacagcggtt cattggtcta tttccgcaag atcccggtgg
     3121 gaaaagtcta cgactatgcc atcaatccca acaagcaagg cgtggtgatt gatgtcctga
     3181 tcgagcggcg ttttaccgat ctggtgaaaa aaggtagccg tttctggaac gtttccggcg
     3241 ttgatgccaa cgtcagtatc agtggcgcga aggtgaaact ggaaagcctg gcggcactgg
     3301 ttaacggtgc gattgccttc gattcaccag aagagtcgaa acctgccgag gcggaagata
     3361 cctttggtct gtatgaagat ctggcccaca gccagcgtgg cgtaataata aaactggaac
     3421 tgccgagtgg ggccggatta accgccgact cgacgccgtt aatgtatcag gggctggaag
     3481 tcggacagct gactaaactg gatttaaatc ctggtggtaa agtcacaggg gaaatgaccg
     3541 ttgatcccag cgtcgttacc ctgcttcgtg aaaatacccg catcgaatta cgcaacccga
     3601 aattatccct tagcgatgct aatctcagcg ccctgctgac cggcaaaacc ttcgagctgg
     3661 tgcccggcga tggcgagcca cgcaaagagt tcgttgttgt gccaggcgaa aaagcactgc
     3721 tgcatgaacc tgatgttctg acgctgaccc tgaccgcgcc ggaaagttac ggtattgatg
     3781 cgggtcagcc gctcattctt cacggcgtgc aggtaggcca ggttatcgat cgtaaactca
     3841 ccagcaaagg cgtcaccttt accgtcgcca tcgagcctca gcatcgcgaa ctggtaaaag
     3901 gcgatagcaa atttgtcgtc aacagccgtg tcgatgtgaa ggtggggctg gatggcgttg
     3961 agtttctcgg tgccagcgcc tcagaatgga tcaatggcgg gatacgtatt ctgccgggcg
     4021 ataaaggtga gatgaaagcc agctatccac tgtatgccaa tttggaaaaa gcgctggaga
     4081 acagccttag cgatttaccc accacaaccg tgagtttgag tgcagagacg ctgccggatg
     4141 tgcaggcagg atcggtagtg ctgtaccgta aatttgaagt tggtgaagtg attaccgtgc
     4201 gtccgcgagc taacgcgttt gatatcgatc tgcatattaa gccggagtat cgcaaccttc
     4261 tgaccagcaa tagcgtgttc tgggcagaag gcggggcgaa agttcagctg aatggtagtg
     4321 gtctgaccgt acaggcatcc ccgctctcca gagcattaaa gggagccatt agcttcgaca
     4381 acctcagcgg tgccagcgcc agtcagcgta aaggcgataa gcgtattctg tatgcttccg
     4441 aaacagcggc ccgtgcggtt ggcgggcaga ttacgcttca cgctttcgat gccggaaaac
     4501 tggcggtcgg gatgccaatt cgctatctcg gtattgatat cgggcaaatc cagacgctgg
     4561 atctgattac cgcacgcaat gaagtacagg caaaggcggt actttacccg gaatatgtcc
     4621 agacctttgc tcgcggtggt acgcgcttct cagtggtcac accgcaaatt tcggcaaatg
     4681 gcgttgagca tcttgatact atcctccagc cgtatatcaa cgtcgaacca ggccggggca
     4741 atcctcgccg cgactttgaa ttacaagagg ccaccattac tgattcgcgt tacctggatg
     4801 gcttaagcat tattgttgaa gcgccggaag ccggttcgtt aggcatcggt acgcctgtgc
     4861 tgttccgtgg tctggaagtc ggtacggtta caggaatgac gctggggaca ttgtcagatc
     4921 gcgtgatgat tgcgatgcgc atcagtaaac gctatcaaca cctggtgcgt aacaattccg
     4981 tcttctggtt ggcatcgggt tacagtctgg actttggtct gacgggcggc gtagtgaaaa
     5041 ccggcacctt taaccagttt atccgtggcg gcatcgcctt cgccacgcct ccgggtacgc
     5101 cactggcacc gaaagcccag gaaggcaaac acttcctgtt gcaggaaagt gaaccgaaag
     5161 agtggcgtga atggggaact gcgcttccca aataatgccc acctcgagca gatccggctg
     5221 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     5281 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     5341 ccggataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5401 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5461 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5521 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5581 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5641 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5701 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5761 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     5821 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5881 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5941 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6001 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6061 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     6121 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6181 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6241 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6301 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6361 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6421 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6721 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6961 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tcc
//
