LOCUS       sequence-334319-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant A598N.
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: pBEL1867
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..97
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(91..282)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     791..810
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        791..809
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             841..863
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4821..4839)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4835..4882
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4927..4945)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5013..5117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5118..5186,5187..5978)
                     /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(5336..5355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6149..6737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6638..6657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6891..6908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6923..7062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6988..7007)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
       61 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      121 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
      181 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
      241 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
      301 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
      361 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
      421 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
      481 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
      541 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
      601 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
      661 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
      721 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatctcgat
      781 cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc tagaaataat
      841 tttgtttaac tttaagaagg agatatacat atggctctta acacaccaca aattacgccg
      901 acaaaaaaga taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt
      961 tgcccgcaat gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat
     1021 tgtccgcgct gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg
     1081 gcaatggcct tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat
     1141 atctggctgt taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc
     1201 aaacagggcg atgcgataac ggggtcgatg gtctttttct gcgttattgg tgccccgctc
     1261 attctggtga cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt
     1321 ccggtactgc tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc
     1381 ggcattggcg ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc
     1441 ttgttctctt ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc
     1501 gaagaactgt gggagcgatt ttatccgcag cgccccgcta cgcgtaggga cgagaaactt
     1561 cgtgtctgcc tagggtgcca ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt
     1621 tgccatatcc cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg
     1681 ttagcgtcaa tcgttttatt gttacctgcc aacctgttgc ctatttctat catttatctg
     1741 aacggaggac ggcaggaaga tacaattctt tccggaatta tgtcgctggc aagtagcaac
     1801 attgcggttg caggaatcgt gtttatcgcc agtattctgg tcccgtttac taaagtgatc
     1861 gtcatgttca ctttactgtt gagcattcat tttaaatgcc agcaaggttt acgcacacgc
     1921 attctgttac tgcggatggt gacctggatt ggtcgctggt cgatgctcga cctgtttgtc
     1981 atatctttaa ccatgtcgct gattaatcgc gatcagatcc tcgcttttac tatgggaccg
     2041 gctgcgtttt atttcggcgc agcggtaatt ttgactattc ttgctgtgga atggctggac
     2101 agccgcttac tttgggatgc acatgagtca ggaaacgccc gcttcgacga ctgaagcgca
     2161 gattaaaaat aaacgccgta tctcaccttt ctggctgctg ccgttcatcg cgctaatgat
     2221 tgccagttgg ctgatttggg acagttatca ggaccggggt aataccgtca ccatcgactt
     2281 tatgtcggcg gatggtattg ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt
     2341 cggaacagtg caggacatca gcctcagcga cgatcttcgt aagattgaag tcaaggtcag
     2401 catcaagtcc gatatgaaag atgcgctgcg cgaagagact cagttctggc tggtgacgcc
     2461 aaaagcatcg ttggcaggtg tctccgggct ggacgccctc gtcggtggga actatatcgg
     2521 catgatgccc gggaaaggta aagagcagga tcactttgtc gcactcgata cccaaccgaa
     2581 atatcggctg gacaatggcg atctgatgat ccacctgcaa gcccccgatc tcggttcgct
     2641 gaacagcggt tcattggtct atttccgcaa gatcccggtg ggaaaagtct acgactatgc
     2701 catcaatccc aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc gttttaccga
     2761 tctggtgaaa aaaggtagcc gtttctggaa cgtttccggc gttgatgcca acgtcagtat
     2821 cagtggcgcg aaggtgaaac tggaaagctt ggcggcactg gttaacggtg cgattgcctt
     2881 cgattcacca gaagagtcga aacctgccga ggcggaagat acctttggtc tgtatgaaga
     2941 tctggcccac agccagcgtg gcgtaataat aaaactggaa ctgccgagtg gggccggatt
     3001 aaccgccgac tcgacgccgt taatgtatca ggggctggaa gtcggacagc tgactaaact
     3061 ggatttaaat cctggtggta aagtcacagg ggaaatgacc gttgatccca gcgtcgttac
     3121 cctgcttcgt gaaaataccc gcatcgaatt acgcaacccg aaattatccc ttagcgatgc
     3181 taatctcagc gccctgctga ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc
     3241 acgcaaagaa ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac ctgatgttct
     3301 gacgctgacc ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc cgctcattct
     3361 tcacggcgtg caggtaggcc aggttatcga tcgtaaactc accagcaaag gcgtcacctt
     3421 taccgtcgcc atcgagcctc agcatcgcga actggtaaaa ggcgatagca aatttgtcgt
     3481 caacagccgt gtcgatgtga aggtggggct ggatggcgtt gagtttctcg gtgccagcgc
     3541 ctcagaatgg atcaatggcg ggatacgtat tctgccgggc gataaaggtg agatgaaagc
     3601 cagctatcca ctgtatgcca atttggaaaa agcgctggag aacagcctta gcgatttacc
     3661 caccacaacc gtgagtttga gtgcagagac gctgccggat gtgcaggcag gatcggtagt
     3721 gctgtaccgt aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt
     3781 tgatatcgat ctgcatatta agccggagta tcgcaacctt ctgaccagca atagcgtgtt
     3841 ctgggcagaa ggcggggcga aagttcagct gaatggtagt ggtctgaccg tacaggcatc
     3901 cccgctctcc agaaacttaa agggagccat tagcttcgac aacctcagcg gtgccagcgc
     3961 cagtcagcgt aaaggcgata agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt
     4021 tggcgggcag attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat
     4081 tcgctatctc ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa
     4141 tgaagtacag gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg
     4201 tacgcgcttc tcagtggtca caccgcaaat ttcggcagct ggcgttgagc atcttgatac
     4261 tatcctccag ccgtatatca acgtcgaacc aggccggggc aatcctcgcc gcgactttga
     4321 attacaagag gccaccatta ctgattcgcg ttacctggat ggcttaagca ttattgttga
     4381 agcgccggaa gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt
     4441 cggtacggtt acaggaatga cgctggggac attgtcagat cgcgtgatga ttgcgatgcg
     4501 catcagtaaa cgctatcaac acctggtgcg taacaattcc gtcttctggt tggcatcggg
     4561 ttacagtctg gactttggtc tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt
     4621 tatccgtggc ggcatcgcct tcgccacgcc tccgggtacg ccactggcac cgaaagccca
     4681 ggaaggcaaa cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac
     4741 tgcgcttccc aaataatgcc cacctcgagc agatccggct gctaacaaag cccgaaagga
     4801 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     4861 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac
     4921 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4981 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5041 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5101 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5161 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5221 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5281 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5341 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5401 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5461 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5521 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5581 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5641 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5701 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5761 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5821 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5881 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5941 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6001 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6061 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6121 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6241 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6601 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6661 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6721 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6781 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6841 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6901 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6961 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7021 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca ccc
//
