LOCUS       sequence-334343-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant V126N.
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: pBEL2345
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4031..4049)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4045..4092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4137..4155)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4223..4327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4328..4396,4397..5188)
                     /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(4546..4565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5359..5947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5848..5867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6101..6118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6133..6272
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6198..6217)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6358..6380
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6374..6565)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atggctctta acacaccaca aattacgccg acaaaaaaga
      121 taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt tgcccgcaat
      181 gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat tgtccgcgct
      241 gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg gcaatggcct
      301 tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat atctggctgt
      361 taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc aaacagggcg
      421 atgcgataac ggggtcgatg gtctttttct gcgttattgg tgccccgctc attctggtga
      481 cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt ccggtactgc
      541 tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc ggcattggcg
      601 ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc ttgttctctt
      661 ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc gaagaactgt
      721 gggagcgatt ttatccgcag cgccccgcta cgcgtaggga cgagaaactt cgtgtctgtc
      781 ttgggtgcca ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt tgccatatcc
      841 cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg ttagcgtcaa
      901 tcgttttatt gttacctgcc aacctgttgc ctatttctat catttatctg aacggaggac
      961 ggcaggaaga tacaattctt tccggaatta tgtcgctggc aagtagcaac attgcggttg
     1021 caggaatcgt gtttatcgcc agtattctgg taccgtttac taaagtgatc gtcatgttca
     1081 ctttactgtt gagcattcat tttaaatgcc agcaaggttt acgcacacgc attctgttac
     1141 tgcggatggt gacctggatt ggtcgctggt cgatgctcga cctgtttgtc atatctttaa
     1201 ccatgtcgct gattaatcgc gatcagatcc tcgcttttac tatgggaccg gctgcgtttt
     1261 atttcggcgc agcggtaatt ttgactattc ttgctgtgga atggctggac agccgcttac
     1321 tttgggatgc acatgagtca ggaaacgccc gcttcgacga ctgaagcgca gattaaaaat
     1381 aaacgccgta tctcaccttt ctggctgctg ccgttcatcg cgctaatgat tgccagttgg
     1441 ctgatttggg acagttatca ggaccggggt aataccgtca ccatcgactt tatgtcggcg
     1501 gatggtattg ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt cggaacagtg
     1561 caggatatca gcctcagcga cgatcttcgt aagattgaag tcaaggtcag catcaagtcc
     1621 gatatgaaag atgcgctgcg cgaagagact cagttctggc tggtgacgcc aaaagcatcg
     1681 ttggcaggtg tctccgggct ggacgccctc aatggtggga actatatcgg catgatgccg
     1741 ggtaaaggta aagagcagga tcactttgtc gcactcgata cccaaccgaa atatcggctg
     1801 gacaatggcg atctgatgat ccacctgcaa gcccccgatc tcggttcgct gaacagcggt
     1861 tcattggtct atttccgcaa gatcccggtg ggaaaagtct acgactatgc catcaatccc
     1921 aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc gttttaccga tctggtgaaa
     1981 aaaggtagcc gtttctggaa cgtttccggc gttgatgcca acgtcagtat cagtggcgcg
     2041 aaggtgaaac tggaaagcct ggcggcactg gttaacggtg cgattgcctt cgattcacca
     2101 gaagagtcga aacctgccga ggcggaagat acctttggtc tgtatgaaga tctggcccac
     2161 agccagcgtg gcgtaataat aaaactggaa ctgccgagtg gggccggatt aaccgccgac
     2221 tcgacgccgt taatgtatca ggggctggaa gtcggacagc tgactaaact ggatttaaat
     2281 cctggtggta aagtcacagg ggaaatgacc gttgatccca gcgtcgttac cctgcttcgt
     2341 gaaaataccc gcatcgaatt acgcaacccg aaattatccc ttagcgatgc taatctcagc
     2401 gccctgctga ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc acgcaaagag
     2461 ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac ctgatgttct gacgctgacc
     2521 ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc cgctcattct tcacggcgtg
     2581 caggtaggcc aggttatcga tcgtaaactc accagcaaag gcgtcacctt taccgtcgcc
     2641 atcgagcctc agcatcgcga actggtaaaa ggcgatagca aatttgtcgt caacagccgt
     2701 gtcgatgtga aggtggggct ggatggcgtt gagtttctcg gtgccagcgc ctcagaatgg
     2761 atcaatggcg ggatacgtat tctgccgggc gataaaggtg agatgaaagc cagctatcca
     2821 ctgtatgcca atttggaaaa agcgctggag aacagcctta gcgatttacc caccacaacc
     2881 gtgagtttga gtgcagagac gctgccggat gtgcaggcag gatcggtagt gctgtaccgt
     2941 aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt tgatatcgat
     3001 ctgcatatta agccggagta tcgcaacctt ctgaccagca atagcgtgtt ctgggcagaa
     3061 ggcggggcga aagttcagct gaatggtagt ggtctgaccg tacaggcatc cccgctctcc
     3121 agagcattaa agggagccat tagcttcgac aacctcagcg gtgccagcgc cagtcagcgt
     3181 aaaggcgata agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt tggcgggcag
     3241 attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat tcgctatctc
     3301 ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa tgaagtacag
     3361 gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg tacgcgcttc
     3421 tcagtggtca caccgcaaat ttcggcagct ggcgttgagc atcttgatac tatcctccag
     3481 ccgtatatca acgtcgaacc aggccggggc aatcctcgcc gcgactttga attacaagag
     3541 gccaccatta ctgattcgcg ttacctggat ggcttaagca ttattgttga agcgccggaa
     3601 gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt cggtacggtt
     3661 acaggaatga cgctggggac attgtcagat cgcgtgatga ttgcgatgcg catcagtaaa
     3721 cgctatcaac acctggtgcg taacaattcc gtcttctggt tggcatcggg ttacagtctg
     3781 gactttggtc tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt tatccgtggc
     3841 ggcatcgcct tcgccacgcc tccgggtacg ccactggcac cgaaagccca ggaaggcaaa
     3901 cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac tgcgcttccc
     3961 aaataatgcc cacctcgagc agatccggct gctaacaaag cccgaaagga agctgagttg
     4021 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4081 aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac gaaagggcct
     4141 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4201 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4261 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4321 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4381 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4441 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4501 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4561 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4621 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4681 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4741 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4801 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4861 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4921 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4981 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5041 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5101 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5161 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5221 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5281 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5341 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5401 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5461 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5521 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5581 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5641 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5701 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5761 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5821 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5881 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5941 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6001 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6061 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6121 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6181 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     6241 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     6301 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6361 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     6421 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     6481 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     6541 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     6601 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     6661 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     6721 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     6781 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     6841 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     6901 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     6961 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     7021 tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga aat
//
