LOCUS       sequence-333960-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant W476N.
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: pBEL1864
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     285..302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    317..456
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(382..401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     542..564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(558..749)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1258..1277
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1258..1276
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1308..1330
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5288..5306)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5302..5349
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5394..5412)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5480..5584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5585..5653,5654..6445)
                     /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(5803..5822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6616..7073,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      301 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
      361 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
      421 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
      481 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
      541 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
      601 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
      661 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
      721 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
      781 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
      841 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
      901 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
      961 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     1021 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     1081 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     1141 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     1201 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa
     1261 tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt
     1321 aagaaggaga tatacatatg gctcttaaca caccacaaat tacgccgaca aaaaagataa
     1381 cagtgagggc aatcggcgag gaactgccgc gtggtgatta ccaacgttgc ccgcaatgtg
     1441 acatgctgtt tagcctgccc gagataaatt ctcatcaaag tgcctattgt ccgcgctgtc
     1501 aggcaaaaat tcgtgacggg cgcgactggt cgctaacgcg cctggcggca atggccttca
     1561 ccatgctgtt gcttatgccg tttgcctggg gcgaaccgct gttgcatatc tggctgttag
     1621 gcattcgtat tgacgccaac gttatgcaag gcatctggca aatgaccaaa cagggcgatg
     1681 cgataacggg gtcgatggtc tttttctgcg ttattggtgc cccgctcatt ctggtgacct
     1741 ccatagctta tttatggttt ggtaaccgac tgggaatgaa tttacgtccg gtactgctga
     1801 tgcttgagcg acttaaagag tgggtaatgc tcgatatcta cctggtcggc attggcgttg
     1861 cttctataaa ggtacaggat tatgcccata tccaggctgg tgtcggcttg ttctcttttg
     1921 tggcgttggt gattttaacg acggtgacgt tgtcacatct taatgtcgaa gaactgtggg
     1981 agcgatttta tccgcagcgc cccgctacgc gtagggacga gaaacttcgt gtctgcctag
     2041 ggtgccattt taccggctat ccagatcagc gtggtcgctg cccgcgttgc catatcccgc
     2101 tacgcctgcg tcgccgtcat agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg
     2161 ttttattgtt acctgccaac ctgttgccta tttctatcat ttatctgaac ggaggacggc
     2221 aggaagatac aattctttcc ggaattatgt cgctggcaag tagcaacatt gcggttgcag
     2281 gaatcgtgtt tatcgccagt attctggtcc cgtttactaa agtgatcgtc atgttcactt
     2341 tactgttgag cattcatttt aaatgccagc aaggtttacg cacacgcatt ctgttactgc
     2401 ggatggtgac ctggattggt cgctggtcga tgctcgacct gtttgtcata tctttaacca
     2461 tgtcgctgat taatcgcgat cagatcctcg cttttactat gggaccggct gcgttttatt
     2521 tcggcgcagc ggtaattttg actattcttg ctgtggaatg gctggacagc cgcttacttt
     2581 gggatgcaca tgagtcagga aacgcccgct tcgacgactg aagcgcagat taaaaataaa
     2641 cgccgtatct cacctttctg gctgctgccg ttcatcgcgc taatgattgc cagttggctg
     2701 atttgggaca gttatcagga ccggggtaat accgtcacca tcgactttat gtcggcggat
     2761 ggtattgttc caggccgtac gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag
     2821 gacatcagcc tcagcgacga tcttcgtaag attgaagtca aggtcagcat caagtccgat
     2881 atgaaagatg cgctgcgcga agagactcag ttctggctgg tgacgccaaa agcatcgttg
     2941 gcaggtgtct ccgggctgga cgccctcgtc ggtgggaact atatcggcat gatgccgggt
     3001 aaaggtaaag agcaggatca ctttgtcgca ctcgataccc aaccgaaata tcggctggac
     3061 aatggcgatc tgatgatcca cctgcaagcc cccgatctcg gttcgctgaa cagcggttca
     3121 ttggtctatt tccgcaagat cccggtggga aaagtctacg actatgccat caatcccaac
     3181 aagcaaggcg tggtgattga tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa
     3241 ggtagccgtt tctggaacgt ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag
     3301 gtgaaactgg aaagcctggc ggcactggtt aacggtgcga ttgccttcga ttcaccagaa
     3361 gagtcgaaac ctgccgaggc ggaagatacc tttggtctgt atgaagatct ggcccacagc
     3421 cagcgtggcg taataataaa actggaactg ccgagtgggg ccggattaac cgccgactcg
     3481 acgccgttaa tgtatcaggg gctggaagtc ggacagctga ctaaactgga tttaaatcct
     3541 ggtggtaaag tcacagggga aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa
     3601 aatacccgca tcgaattacg caacccgaaa ttatccctta gcgatgctaa tctcagcgcc
     3661 ctgctgaccg gcaaaacctt cgagctggtg cccggcgatg gcgagccacg caaagagttc
     3721 gttgttgtgc caggcgaaaa agcactgctg catgaacctg atgttctgac gctgaccctg
     3781 accgcgccgg aaagttacgg tattgatgcg ggtcagccgc tcattcttca cggcgtgcag
     3841 gtaggccagg ttatcgatcg taaactcacc agcaaaggcg tcacctttac cgtcgccatc
     3901 gagcctcagc atcgcgaact ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc
     3961 gatgtgaagg tggggctgga tggcgttgag tttctcggtg ccagcgcctc agaaaacatc
     4021 aatggcggga tacgtattct gccgggcgat aaaggtgaga tgaaagccag ctatccactg
     4081 tatgccaatt tggaaaaagc gctggagaac agccttagcg atttacccac cacaaccgtg
     4141 agtttgagtg cagagacgct gccggatgtg caggcaggat cggtagtgct gtaccgtaaa
     4201 tttgaagttg gtgaagtgat taccgtgcgt ccgcgagcta acgcgtttga catcgatctg
     4261 catattaagc cggagtatcg caaccttctg accagcaata gcgtgttctg ggcagaaggc
     4321 ggggcgaaag ttcagctgaa tggtagtggt ctgaccgtac aggcatcccc gctctccaga
     4381 gctctaaagg gagccattag cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa
     4441 ggcgataagc gtattctgta tgcttccgaa acagcggccc gtgcggttgg cgggcagatt
     4501 acgcttcacg ctttcgatgc cggaaaactg gcggtcggga tgccaattcg ctatctcggt
     4561 attgacatcg ggcaaatcca gacgctggat ctgattaccg cacgcaatga agtacaggca
     4621 aaggcggtac tttacccgga atatgtccag acctttgctc gcggtggtac gcgcttctca
     4681 gtggtcacac cgcaaatttc ggcagctggc gttgagcatc ttgatactat cctccagccg
     4741 tatatcaacg tcgaaccagg ccggggcaat cctcgccgcg actttgaatt acaagaggcc
     4801 accattactg attcgcggta cctggatggc ttaagcatta ttgttgaagc gccggaagcc
     4861 ggttcgttag gcatcggtac gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca
     4921 ggaatgacgc tggggacatt gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc
     4981 tatcaacacc tggtgcgtaa caattccgtc ttctggttgg catcgggtta cagtctggac
     5041 tttggtctga cgggcggcgt agtgaaaacc ggcaccttta accagtttat ccgtggcggc
     5101 atcgccttcg ccacgcctcc gggtacgcca ctggcaccga aagcccagga aggcaaacac
     5161 ttcctgttgc aggaaagtga accgaaagag tggcgtgaat ggggaactgc gcttcccaaa
     5221 taatgcccac ctcgagcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     5281 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     5341 ggttttttgc tgaaaggagg aactatatcc ggataattct tgaagacgaa agggcctcgt
     5401 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5581 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5641 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5701 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5761 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5821 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5881 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5941 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6001 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6061 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6121 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6181 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6241 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6301 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6361 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6421 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6481 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6541 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6601 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6661 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6721 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6781 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6841 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6901 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6961 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7021 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcg
//
