LOCUS       sequence-334341-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant L125N.
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: pBEL2344
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..87
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(81..272)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     781..800
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        781..799
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             831..853
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4811..4829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4825..4872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4917..4935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5003..5107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5108..5176,5177..5968)
                     /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(5326..5345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6139..6727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6628..6647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6881..6898
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6913..7052
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6978..6997)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
       61 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
      121 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
      181 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
      241 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
      301 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
      361 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
      421 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
      481 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
      541 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
      601 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
      661 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
      721 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatctcgat cccgcgaaat
      781 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
      841 tttaagaagg agatatacat atggctctta acacaccaca aattacgccg acaaaaaaga
      901 taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt tgcccgcaat
      961 gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat tgtccgcgct
     1021 gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg gcaatggcct
     1081 tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat atctggctgt
     1141 taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc aaacagggcg
     1201 atgcgataac ggggtcgatg gtctttttct gcgttattgg tgccccgctc attctggtga
     1261 cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt ccggtactgc
     1321 tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc ggcattggcg
     1381 ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc ttgttctctt
     1441 ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc gaagaactgt
     1501 gggagcgatt ttatccgcag cgccccgcta cgcgtaggga cgagaaactt cgtgtctgtc
     1561 ttgggtgcca ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt tgccatatcc
     1621 cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg ttagcgtcaa
     1681 tcgttttatt gttacctgcc aacctgttgc ctatttctat catttatctg aacggaggac
     1741 ggcaggaaga tacaattctt tccggaatta tgtcgctggc aagtagcaac attgcggttg
     1801 caggaatcgt gtttatcgcc agtattctgg taccgtttac taaagtgatc gtcatgttca
     1861 ctttactgtt gagcattcat tttaaatgcc agcaaggttt acgcacacgc attctgttac
     1921 tgcggatggt gacctggatt ggtcgctggt cgatgctcga cctgtttgtc atatctttaa
     1981 ccatgtcgct gattaatcgc gatcagatcc tcgcttttac tatgggaccg gctgcgtttt
     2041 atttcggcgc agcggtaatt ttgactattc ttgctgtgga atggctggac agccgcttac
     2101 tttgggatgc acatgagtca ggaaacgccc gcttcgacga ctgaagcgca gattaaaaat
     2161 aaacgccgta tctcaccttt ctggctgctg ccgttcatcg cgctaatgat tgccagttgg
     2221 ctgatttggg acagttatca ggaccggggt aataccgtca ccatcgactt tatgtcggcg
     2281 gatggtattg ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt cggaacagtg
     2341 caggatatca gcctcagcga cgatcttcgt aagattgaag tcaaggtcag catcaagtcc
     2401 gatatgaaag atgcgctgcg cgaagagact cagttctggc tggtgacgcc aaaagcatcg
     2461 ttggcaggtg tctccgggct ggacgccaat gtcggtggga actatatcgg catgatgccg
     2521 ggtaaaggta aagagcagga tcactttgtc gcactcgata cccaaccgaa atatcggctg
     2581 gacaatggcg atctgatgat ccacctgcaa gcccccgatc tcggttcgct gaacagcggt
     2641 tcattggtct atttccgcaa gatcccggtg ggaaaagtct acgactatgc catcaatccc
     2701 aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc gttttaccga tctggtgaaa
     2761 aaaggtagcc gtttctggaa cgtttccggc gttgatgcca acgtcagtat cagtggcgcg
     2821 aaggtgaaac tggaaagcct ggcggcactg gttaacggtg cgattgcctt cgattcacca
     2881 gaagagtcga aacctgccga ggcggaagat acctttggtc tgtatgaaga tctggcccac
     2941 agccagcgtg gcgtaataat aaaactggaa ctgccgagtg gggccggatt aaccgccgac
     3001 tcgacgccgt taatgtatca ggggctggaa gtcggacagc tgactaaact ggatttaaat
     3061 cctggtggta aagtcacagg ggaaatgacc gttgatccca gcgtcgttac cctgcttcgt
     3121 gaaaataccc gcatcgaatt acgcaacccg aaattatccc ttagcgatgc taatctcagc
     3181 gccctgctga ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc acgcaaagag
     3241 ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac ctgatgttct gacgctgacc
     3301 ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc cgctcattct tcacggcgtg
     3361 caggtaggcc aggttatcga tcgtaaactc accagcaaag gcgtcacctt taccgtcgcc
     3421 atcgagcctc agcatcgcga actggtaaaa ggcgatagca aatttgtcgt caacagccgt
     3481 gtcgatgtga aggtggggct ggatggcgtt gagtttctcg gtgccagcgc ctcagaatgg
     3541 atcaatggcg ggatacgtat tctgccgggc gataaaggtg agatgaaagc cagctatcca
     3601 ctgtatgcca atttggaaaa agcgctggag aacagcctta gcgatttacc caccacaacc
     3661 gtgagtttga gtgcagagac gctgccggat gtgcaggcag gatcggtagt gctgtaccgt
     3721 aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt tgatatcgat
     3781 ctgcatatta agccggagta tcgcaacctt ctgaccagca atagcgtgtt ctgggcagaa
     3841 ggcggggcga aagttcagct gaatggtagt ggtctgaccg tacaggcatc cccgctctcc
     3901 agagcattaa agggagccat tagcttcgac aacctcagcg gtgccagcgc cagtcagcgt
     3961 aaaggcgata agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt tggcgggcag
     4021 attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat tcgctatctc
     4081 ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa tgaagtacag
     4141 gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg tacgcgcttc
     4201 tcagtggtca caccgcaaat ttcggcagct ggcgttgagc atcttgatac tatcctccag
     4261 ccgtatatca acgtcgaacc aggccggggc aatcctcgcc gcgactttga attacaagag
     4321 gccaccatta ctgattcgcg ttacctggat ggcttaagca ttattgttga agcgccggaa
     4381 gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt cggtacggtt
     4441 acaggaatga cgctggggac attgtcagat cgcgtgatga ttgcgatgcg catcagtaaa
     4501 cgctatcaac acctggtgcg taacaattcc gtcttctggt tggcatcggg ttacagtctg
     4561 gactttggtc tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt tatccgtggc
     4621 ggcatcgcct tcgccacgcc tccgggtacg ccactggcac cgaaagccca ggaaggcaaa
     4681 cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac tgcgcttccc
     4741 aaataatgcc cacctcgagc agatccggct gctaacaaag cccgaaagga agctgagttg
     4801 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     4861 aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac gaaagggcct
     4921 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4981 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5101 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5161 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5221 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5281 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5341 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5401 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5461 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5521 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5581 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5641 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5701 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5761 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5821 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5881 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5941 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6001 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6061 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6241 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6781 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6841 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6901 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6961 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     7021 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca ccc
//
