LOCUS       sequence-333956-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant L595N.
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: pBEL1866
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     411..430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     664..681
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    696..835
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(761..780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     921..943
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(937..1128)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1637..1656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1637..1655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1687..1709
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5667..5685)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5681..5728
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5773..5791)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5859..5963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5964..6032,6033..6824)
                     /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(6182..6201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6995..7073,1..510)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
       61 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      121 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      181 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      241 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      301 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      361 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      421 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      481 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      541 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      601 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      661 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
      721 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
      781 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
      841 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      901 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      961 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     1021 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     1081 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     1141 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     1201 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     1261 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     1321 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     1381 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     1441 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     1501 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     1561 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     1621 ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctaga
     1681 aataattttg tttaacttta agaaggagat atacatatgg ctcttaacac accacaaatt
     1741 acgccgacaa aaaagataac agtgagggca atcggcgagg aactgccgcg tggtgattac
     1801 caacgttgcc cgcaatgtga catgctgttt agcctgcccg agataaattc tcatcaaagt
     1861 gcctattgtc cgcgctgtca ggcaaaaatt cgtgacgggc gcgactggtc gctaacgcgc
     1921 ctggcggcaa tggccttcac catgctgttg cttatgccgt ttgcctgggg cgaaccgctg
     1981 ttgcatatct ggctgttagg cattcgtatt gacgccaacg ttatgcaagg catctggcaa
     2041 atgaccaaac agggcgatgc gataacgggg tcgatggtct ttttctgcgt tattggtgcc
     2101 ccgctcattc tggtgacctc catagcttat ttatggtttg gtaaccgact gggaatgaat
     2161 ttacgtccgg tactgctgat gcttgagcga cttaaagagt gggtaatgct cgatatctac
     2221 ctggtcggca ttggcgttgc ttctataaag gtacaggatt atgcccatat ccaggctggt
     2281 gtcggcttgt tctcttttgt ggcgttggtg attttaacga cggtgacgtt gtcacatctt
     2341 aatgtcgaag aactgtggga gcgattttat ccgcagcgcc ccgctacgcg tagggacgag
     2401 aaacttcgtg tctgcctagg gtgccatttt accggctatc cagatcagcg tggtcgctgc
     2461 ccgcgttgcc atatcccgct acgcctgcgt cgccgtcata gtctgcaaaa atgctgggcg
     2521 gcgctgttag cgtcaatcgt tttattgtta cctgccaacc tgttgcctat ttctatcatt
     2581 tatctgaacg gaggacggca ggaagataca attctttccg gaattatgtc gctggcaagt
     2641 agcaacattg cggttgcagg aatcgtgttt atcgccagta ttctggtccc gtttactaaa
     2701 gtgatcgtca tgttcacttt actgttgagc attcatttta aatgccagca aggtttacgc
     2761 acacgcattc tgttactgcg gatggtgacc tggattggtc gctggtcgat gctcgacctg
     2821 tttgtcatat ctttaaccat gtcgctgatt aatcgcgatc agatcctcgc ttttactatg
     2881 ggaccggctg cgttttattt cggcgcagcg gtaattttga ctattcttgc tgtggaatgg
     2941 ctggacagcc gcttactttg ggatgcacat gagtcaggaa acgcccgctt cgacgactga
     3001 agcgcagatt aaaaataaac gccgtatctc acctttctgg ctgctgccgt tcatcgcgct
     3061 aatgattgcc agttggctga tttgggacag ttatcaggac cggggtaata ccgtcaccat
     3121 cgactttatg tcggcggatg gtattgttcc aggccgtacg cctgttcgtt atcagggcgt
     3181 tgaagtcgga acagtgcagg acatcagcct cagcgacgat cttcgtaaga ttgaagtcaa
     3241 ggtcagcatc aagtccgata tgaaagatgc gctgcgcgaa gagactcagt tctggctggt
     3301 gacgccaaaa gcatcgttgg caggtgtctc cgggctggac gccctcgtcg gtgggaacta
     3361 tatcggcatg atgcccggga aaggtaaaga gcaggatcac tttgtcgcac tcgataccca
     3421 accgaaatat cggctggaca atggcgatct gatgatccac ctgcaagccc ccgatctcgg
     3481 ttcgctgaac agcggttcat tggtctattt ccgcaagatc ccggtgggaa aagtctacga
     3541 ctatgccatc aatcccaaca agcaaggcgt ggtgattgat gtcctgatcg agcggcgttt
     3601 taccgatctg gtgaaaaaag gtagccgttt ctggaacgtt tccggcgttg atgccaacgt
     3661 cagtatcagt ggcgcgaagg tgaaactgga aagcttggcg gcactggtta acggtgcgat
     3721 tgccttcgat tcaccagaag agtcgaaacc tgccgaggcg gaagatacct ttggtctgta
     3781 tgaagatctg gcccacagcc agcgtggcgt aataataaaa ctggaactgc cgagtggggc
     3841 cggattaacc gccgactcga cgccgttaat gtatcagggg ctggaagtcg gacagctgac
     3901 taaactggat ttaaatcctg gtggtaaagt cacaggggaa atgaccgttg atcccagcgt
     3961 cgttaccctg cttcgtgaaa atacccgcat cgaattacgc aacccgaaat tatcccttag
     4021 cgatgctaat ctcagcgccc tgctgaccgg caaaaccttc gagctggtgc ccggcgatgg
     4081 cgagccacgc aaagaattcg ttgttgtgcc aggcgaaaaa gcactgctgc atgaacctga
     4141 tgttctgacg ctgaccctga ccgcgccgga aagttacggt attgatgcgg gtcagccgct
     4201 cattcttcac ggcgtgcagg taggccaggt tatcgatcgt aaactcacca gcaaaggcgt
     4261 cacctttacc gtcgccatcg agcctcagca tcgcgaactg gtaaaaggcg atagcaaatt
     4321 tgtcgtcaac agccgtgtcg atgtgaaggt ggggctggat ggcgttgagt ttctcggtgc
     4381 cagcgcctca gaatggatca atggcgggat acgtattctg ccgggcgata aaggtgagat
     4441 gaaagccagc tatccactgt atgccaattt ggaaaaagcg ctggagaaca gccttagcga
     4501 tttacccacc acaaccgtga gtttgagtgc agagacgctg ccggatgtgc aggcaggatc
     4561 ggtagtgctg taccgtaaat ttgaagttgg tgaagtgatt accgtgcgtc cgcgagctaa
     4621 cgcgtttgat atcgatctgc atattaagcc ggagtatcgc aaccttctga ccagcaatag
     4681 cgtgttctgg gcagaaggcg gggcgaaagt tcagctgaat ggtagtggtc tgaccgtaca
     4741 ggcatccccg aattccagag cattaaaggg agccattagc ttcgacaacc tcagcggtgc
     4801 cagcgccagt cagcgtaaag gcgataagcg tattctgtat gcttccgaaa cagcggcccg
     4861 tgcggttggc gggcagatta cgcttcacgc tttcgatgcc ggaaaactgg cggtcgggat
     4921 gccaattcgc tatctcggta ttgatatcgg gcaaatccag acgctggatc tgattaccgc
     4981 acgcaatgaa gtacaggcaa aggcggtact ttacccggaa tatgtccaga cctttgctcg
     5041 cggtggtacg cgcttctcag tggtcacacc gcaaatttcg gcagctggcg ttgagcatct
     5101 tgatactatc ctccagccgt atatcaacgt cgaaccaggc cggggcaatc ctcgccgcga
     5161 ctttgaatta caagaggcca ccattactga ttcgcgttac ctggatggct taagcattat
     5221 tgttgaagcg ccggaagccg gttcgttagg catcggtacg cctgtgctgt tccgtggtct
     5281 ggaagtcggt acggttacag gaatgacgct ggggacattg tcagatcgcg tgatgattgc
     5341 gatgcgcatc agtaaacgct atcaacacct ggtgcgtaac aattccgtct tctggttggc
     5401 atcgggttac agtctggact ttggtctgac gggcggcgta gtgaaaaccg gcacctttaa
     5461 ccagtttatc cgtggcggca tcgccttcgc cacgcctccg ggtacgccac tggcaccgaa
     5521 agcccaggaa ggcaaacact tcctgttgca ggaaagtgaa ccgaaagagt ggcgtgaatg
     5581 gggaactgcg cttcccaaat aatgcccacc tcgagcagat ccggctgcta acaaagcccg
     5641 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     5701 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gataattctt
     5761 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5821 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5881 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5941 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6001 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6061 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6121 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6181 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     6241 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6301 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6361 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6421 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6481 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     6541 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6601 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6661 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6721 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6781 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6841 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6901 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6961 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7021 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccg
//
