LOCUS       sequence-345257-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB(V708N).
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: pBEL2419
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      193..781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     682..701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     935..952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    967..1106
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1032..1051)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1192..1214
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1208..1399)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1908..1927
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1908..1926
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1958..1980
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5938..5956)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5952..5999
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6044..6062)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6130..6234
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6235..6303,6304..7073,1..22)
                     /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(6453..6472)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
       61 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
      121 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
      181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      301 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
      361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      841 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      901 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      961 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     1021 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     1081 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     1141 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1201 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     1261 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     1321 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     1381 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     1441 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     1501 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     1561 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     1621 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     1681 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     1741 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     1801 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     1861 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
     1921 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
     1981 tatacatatg gctcttaaca caccacaaat tacgccgaca aaaaagataa cagtgagggc
     2041 aatcggcgag gaactgccgc gtggtgatta ccaacgttgc ccgcaatgtg acatgctgtt
     2101 tagcctgccc gagataaatt ctcatcaaag tgcctattgt ccgcgctgtc aggcaaaaat
     2161 tcgtgacggg cgcgactggt cgctaacgcg cctggcggca atggccttca ccatgctgtt
     2221 gcttatgccg tttgcctggg gcgaaccgct gttgcatatc tggctgttag gcattcgtat
     2281 tgacgccaac gttatgcaag gcatctggca aatgaccaaa cagggcgatg cgataacggg
     2341 gtcgatggtc tttttctgcg ttattggtgc cccgctcatt ctggtgacct ccatagctta
     2401 tttatggttt ggtaaccgac tgggaatgaa tttacgtccg gtactgctga tgcttgagcg
     2461 acttaaagag tgggtaatgc tcgatatcta cctggtcggc attggcgttg cttctataaa
     2521 ggtacaggat tatgcccata tccaggctgg tgtcggcttg ttctcttttg tggcgttggt
     2581 gattttaacg acggtgacgt tgtcacatct taatgtcgaa gaactgtggg agcgatttta
     2641 tccgcagcgc cccgctacgc gtagggacga gaaacttcgt gtctgtcttg ggtgccattt
     2701 taccggctat ccagatcagc gtggtcgctg cccgcgttgc catatcccgc tacgcctgcg
     2761 tcgccgtcat agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg ttttattgtt
     2821 acctgccaac ctgttgccta tttctatcat ttatctgaac ggaggacggc aggaagatac
     2881 aattctttcc ggaattatgt cgctggcaag tagcaacatt gcggttgcag gaatcgtgtt
     2941 tatcgccagt attctggtac cgtttactaa agtgatcgtc atgttcactt tactgttgag
     3001 cattcatttt aaatgccagc aaggtttacg cacacgcatt ctgttactgc ggatggtgac
     3061 ctggattggt cgctggtcga tgctcgacct gtttgtcata tctttaacca tgtcgctgat
     3121 taatcgcgat cagatcctcg cttttactat gggaccggct gcgttttatt tcggcgcagc
     3181 ggtaattttg actattcttg ctgtggaatg gctggacagc cgcttacttt gggatgcaca
     3241 tgagtcagga aacgcccgct tcgacgactg aagcgcagat taaaaataaa cgccgtatct
     3301 cacctttctg gctgctgccg ttcatcgcgc taatgattgc cagttggctg atttgggaca
     3361 gttatcagga ccggggtaat accgtcacca tcgactttat gtcggcggat ggtattgttc
     3421 caggccgtac gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag gatatcagcc
     3481 tcagcgacga tcttcgtaag attgaagtca aggtcagcat caagtccgat atgaaagatg
     3541 cgctgcgcga agagactcag ttctggctgg tgacgccaaa agcatcgttg gcaggtgtct
     3601 ccgggctgga cgccctcgtc ggtgggaact atatcggcat gatgccgggt aaaggtaaag
     3661 agcaggatca ctttgtcgca ctcgataccc aaccgaaata tcggctggac aatggcgatc
     3721 tgatgatcca cctgcaagcc cccgatctcg gttcgctgaa cagcggttca ttggtctatt
     3781 tccgcaagat cccggtggga aaagtctacg actatgccat caatcccaac aagcaaggcg
     3841 tggtgattga tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa ggtagccgtt
     3901 tctggaacgt ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag gtgaaactgg
     3961 aaagcctggc ggcactggtt aacggtgcga ttgccttcga ttcaccagaa gagtcgaaac
     4021 ctgccgaggc ggaagatacc tttggtctgt atgaagatct ggcccacagc cagcgtggcg
     4081 taataataaa actggaactg ccgagtgggg ccggattaac cgccgactcg acgccgttaa
     4141 tgtatcaggg gctggaagtc ggacagctga ctaaactgga tttaaatcct ggtggtaaag
     4201 tcacagggga aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa aatacccgca
     4261 tcgaattacg caacccgaaa ttatccctta gcgatgctaa tctcagcgcc ctgctgaccg
     4321 gcaaaacctt cgagctggtg cccggcgatg gcgagccacg caaagagttc gttgttgtgc
     4381 caggcgaaaa agcactgctg catgaacctg atgttctgac gctgaccctg accgcgccgg
     4441 aaagttacgg tattgatgcg ggtcagccgc tcattcttca cggcgtgcag gtaggccagg
     4501 ttatcgatcg taaactcacc agcaaaggcg tcacctttac cgtcgccatc gagcctcagc
     4561 atcgcgaact ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc gatgtgaagg
     4621 tggggctgga tggcgttgag tttctcggtg ccagcgcctc agaatggatc aatggcggga
     4681 tacgtattct gccgggcgat aaaggtgaga tgaaagccag ctatccactg tatgccaatt
     4741 tggaaaaagc gctggagaac agccttagcg atttacccac cacaaccgtg agtttgagtg
     4801 cagagacgct gccggatgtg caggcaggat cggtagtgct gtaccgtaaa tttgaagttg
     4861 gtgaagtgat taccgtgcgt ccgcgagcta acgcgtttga tatcgatctg catattaagc
     4921 cggagtatcg caaccttctg accagcaata gcgtgttctg ggcagaaggc ggggcgaaag
     4981 ttcagctgaa tggtagtggt ctgaccgtac aggcatcccc gctctccaga gcattaaagg
     5041 gagccattag cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa ggcgataagc
     5101 gtattctgta tgcttccgaa acagcggccc gtgcggttgg cgggcagatt acgcttcacg
     5161 ctttcgatgc cggaaaactg gcggtcggga tgccaattcg ctatctcggt attgatatcg
     5221 ggcaaatcca gacgctggat ctgattaccg cacgcaatga agtacaggca aaggcggtac
     5281 tttacccgga atatgtccag acctttgctc gcggtggtac gcgcttctca gtggtcacac
     5341 cgcaaatttc ggcagctggc aatgagcatc ttgatactat cctccagccg tatatcaacg
     5401 tcgaaccagg ccggggcaat cctcgccgcg actttgaatt acaagaggcc accattactg
     5461 attcgcgtta cctggatggc ttaagcatta ttgttgaagc gccggaagcc ggttcgttag
     5521 gcatcggtac gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca ggaatgacgc
     5581 tggggacatt gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc tatcaacacc
     5641 tggtgcgtaa caattccgtc ttctggttgg catcgggtta cagtctggac tttggtctga
     5701 cgggcggcgt agtgaaaacc ggcaccttta accagtttat ccgtggcggc atcgccttcg
     5761 ccacgcctcc gggtacgcca ctggcaccga aagcccagga aggcaaacac ttcctgttgc
     5821 aggaaagtga accgaaagag tggcgtgaat ggggaactgc gcttcccaaa taatgcccac
     5881 ctcgagcaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     5941 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     6001 tgaaaggagg aactatatcc ggataattct tgaagacgaa agggcctcgt gatacgccta
     6061 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6121 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6181 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6241 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6301 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6361 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6421 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     6481 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6541 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6601 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6661 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6721 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6781 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6841 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6901 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6961 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7021 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgc
//
