LOCUS       sequence-334328-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant I477N.
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: pBEL1865
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     355..374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     608..625
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    640..779
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(705..724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     865..887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(881..1072)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1581..1600
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1581..1599
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1631..1653
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5611..5629)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5625..5672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5717..5735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5803..5907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5908..5976,5977..6768)
                     /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(6126..6145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6939..7073,1..454)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
       61 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      121 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      181 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      241 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      301 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      361 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      421 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      481 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      541 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      601 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
      661 tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat
      721 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
      781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      901 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
      961 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     1021 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     1081 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     1141 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     1201 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     1261 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     1321 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     1381 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     1441 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     1501 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
     1561 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
     1621 tagaaataat tttgtttaac tttaagaagg agatatacat atggctctta acacaccaca
     1681 aattacgccg acaaaaaaga taacagtgag ggcaatcggc gaggaactgc cgcgtggtga
     1741 ttaccaacgt tgcccgcaat gtgacatgct gtttagcctg cccgagataa attctcatca
     1801 aagtgcctat tgtccgcgct gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac
     1861 gcgcctggcg gcaatggcct tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc
     1921 gctgttgcat atctggctgt taggcattcg tattgacgcc aacgttatgc aaggcatctg
     1981 gcaaatgacc aaacagggcg atgcgataac ggggtcgatg gtctttttct gcgttattgg
     2041 tgccccgctc attctggtga cctccatagc ttatttatgg tttggtaacc gactgggaat
     2101 gaatttacgt ccggtactgc tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat
     2161 ctacctggtc ggcattggcg ttgcttctat aaaggtacag gattatgccc atatccaggc
     2221 tggtgtcggc ttgttctctt ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca
     2281 tcttaatgtc gaagaactgt gggagcgatt ttatccgcag cgccccgcta cgcgtaggga
     2341 cgagaaactt cgtgtctgcc tagggtgcca ttttaccggc tatccagatc agcgtggtcg
     2401 ctgcccgcgt tgccatatcc cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg
     2461 ggcggcgctg ttagcgtcaa tcgttttatt gttacctgcc aacctgttgc ctatttctat
     2521 catttatctg aacggaggac ggcaggaaga tacaattctt tccggaatta tgtcgctggc
     2581 aagtagcaac attgcggttg caggaatcgt gtttatcgcc agtattctgg tcccgtttac
     2641 taaagtgatc gtcatgttca ctttactgtt gagcattcat tttaaatgcc agcaaggttt
     2701 acgcacacgc attctgttac tgcggatggt gacctggatt ggtcgctggt cgatgctcga
     2761 cctgtttgtc atatctttaa ccatgtcgct gattaatcgc gatcagatcc tcgcttttac
     2821 tatgggaccg gctgcgtttt atttcggcgc agcggtaatt ttgactattc ttgctgtgga
     2881 atggctggac agccgcttac tttgggatgc acatgagtca ggaaacgccc gcttcgacga
     2941 ctgaagcgca gattaaaaat aaacgccgta tctcaccttt ctggctgctg ccgttcatcg
     3001 cgctaatgat tgccagttgg ctgatttggg acagttatca ggaccggggt aataccgtca
     3061 ccatcgactt tatgtcggcg gatggtattg ttccaggccg tacgcctgtt cgttatcagg
     3121 gcgttgaagt cggaacagtg caggacatca gcctcagcga cgatcttcgt aagattgaag
     3181 tcaaggtcag catcaagtcc gatatgaaag atgcgctgcg cgaagagact cagttctggc
     3241 tggtgacgcc aaaagcatcg ttggcaggtg tctccgggct ggacgccctc gtcggtggga
     3301 actatatcgg catgatgccg ggtaaaggta aagagcagga tcactttgtc gcactcgata
     3361 cccaaccgaa atatcggctg gacaatggcg atctgatgat ccacctgcaa gcccccgatc
     3421 tcggttcgct gaacagcggt tcattggtct atttccgcaa gatcccggtg ggaaaagtct
     3481 acgactatgc catcaatccc aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc
     3541 gttttaccga tctggtgaaa aaaggtagcc gtttctggaa cgtttccggc gttgatgcca
     3601 acgtcagtat cagtggcgcg aaggtgaaac tggaaagcct ggcggcactg gttaacggtg
     3661 cgattgcctt cgattcacca gaagagtcga aacctgccga ggcggaagat acctttggtc
     3721 tgtatgaaga tctggcccac agccagcgtg gcgtaataat aaaactggaa ctgccgagtg
     3781 gggccggatt aaccgccgac tcgacgccgt taatgtatca ggggctggaa gtcggacagc
     3841 tgactaaact ggatttaaat cctggtggta aagtcacagg ggaaatgacc gttgatccca
     3901 gcgtcgttac cctgcttcgt gaaaataccc gcatcgaatt acgcaacccg aaattatccc
     3961 ttagcgatgc taatctcagc gccctgctga ccggcaaaac cttcgagctg gtgcccggcg
     4021 atggcgagcc acgcaaagag ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac
     4081 ctgatgttct gacgctgacc ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc
     4141 cgctcattct tcacggcgtg caggtaggcc aggttatcga tcgtaaactc accagcaaag
     4201 gcgtcacctt taccgtcgcc atcgagcctc agcatcgcga actggtaaaa ggcgatagca
     4261 aatttgtcgt caacagccgt gtcgatgtga aggtggggct ggatggcgtt gagtttctcg
     4321 gtgccagcgc ctcagaatgg aataatggcg ggatacgtat tctgccgggc gataaaggtg
     4381 agatgaaagc cagctatcca ctgtatgcca atttggaaaa agcgctggag aacagcctta
     4441 gcgatttacc caccacaacc gtgagtttga gtgcagagac gctgccggat gtgcaggcag
     4501 gatcggtagt gctgtaccgt aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag
     4561 ctaacgcgtt tgacatcgat ctgcatatta agccggagta tcgcaacctt ctgaccagca
     4621 atagcgtgtt ctgggcagaa ggcggggcga aagttcagct gaatggtagt ggtctgaccg
     4681 tacaggcatc cccgctctcc agagctctaa agggagccat tagcttcgac aacctcagcg
     4741 gtgccagcgc cagtcagcgt aaaggcgata agcgtattct gtatgcttcc gaaacagcgg
     4801 cccgtgcggt tggcgggcag attacgcttc acgctttcga tgccggaaaa ctggcggtcg
     4861 ggatgccaat tcgctatctc ggtattgaca tcgggcaaat ccagacgctg gatctgatta
     4921 ccgcacgcaa tgaagtacag gcaaaggcgg tactttaccc ggaatatgtc cagacctttg
     4981 ctcgcggtgg tacgcgcttc tcagtggtca caccgcaaat ttcggcagct ggcgttgagc
     5041 atcttgatac tatcctccag ccgtatatca acgtcgaacc aggccggggc aatcctcgcc
     5101 gcgactttga attacaagag gccaccatta ctgattcgcg gtacctggat ggcttaagca
     5161 ttattgttga agcgccggaa gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg
     5221 gtctggaagt cggtacggtt acaggaatga cgctggggac attgtcagat cgcgtgatga
     5281 ttgcgatgcg catcagtaaa cgctatcaac acctggtgcg taacaattcc gtcttctggt
     5341 tggcatcggg ttacagtctg gactttggtc tgacgggcgg cgtagtgaaa accggcacct
     5401 ttaaccagtt tatccgtggc ggcatcgcct tcgccacgcc tccgggtacg ccactggcac
     5461 cgaaagccca ggaaggcaaa cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg
     5521 aatggggaac tgcgcttccc aaataatgcc cacctcgagc agatccggct gctaacaaag
     5581 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5641 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataat
     5701 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5761 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5821 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5881 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5941 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6001 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6061 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6121 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     6181 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6241 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6301 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6361 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6421 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     6481 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6541 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6601 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6661 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6721 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6781 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6841 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6901 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6961 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7021 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gat
//
