LOCUS       sequence-334352-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant L711N.
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: pBEL1869
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..55
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    70..209
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(135..154)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     295..317
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(311..502)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1011..1030
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1011..1029
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1061..1083
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5041..5059)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5055..5102
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5147..5165)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5233..5337
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5338..5406,5407..6198)
                     /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(5556..5575)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6369..6957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6858..6877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
       61 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
      121 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
      181 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
      241 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
      301 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
      361 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
      421 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
      481 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
      541 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
      601 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
      661 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
      721 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
      781 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
      841 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
      901 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
      961 gcacccgtgg ccaggaccca acgctgcccg agatctcgat cccgcgaaat taatacgact
     1021 cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg
     1081 agatatacat atggctctta acacaccaca aattacgccg acaaaaaaga taacagtgag
     1141 ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt tgcccgcaat gtgacatgct
     1201 gtttagcctg cccgagataa attctcatca aagtgcctat tgtccgcgct gtcaggcaaa
     1261 aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg gcaatggcct tcaccatgct
     1321 gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat atctggctgt taggcattcg
     1381 tattgacgcc aacgttatgc aaggcatctg gcaaatgacc aaacagggcg atgcgataac
     1441 ggggtcgatg gtctttttct gcgttattgg tgccccgctc attctggtga cctccatagc
     1501 ttatttatgg tttggtaacc gactgggaat gaatttacgt ccggtactgc tgatgcttga
     1561 gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc ggcattggcg ttgcttctat
     1621 aaaggtacag gattatgccc atatccaggc tggtgtcggc ttgttctctt ttgtggcgtt
     1681 ggtgatttta acgacggtga cgttgtcaca tcttaatgtc gaagaactgt gggagcgatt
     1741 ttatccgcag cgccccgcta cgcgtaggga cgagaaactt cgtgtctgcc tagggtgcca
     1801 ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt tgccatatcc cgctacgcct
     1861 gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg ttagcgtcaa tcgttttatt
     1921 gttacctgcc aacctgttgc ctatttctat catttatctg aacggaggac ggcaggaaga
     1981 tacaattctt tccggaatta tgtcgctggc aagtagcaac attgcggttg caggaatcgt
     2041 gtttatcgcc agtattctgg tcccgtttac taaagtgatc gtcatgttca ctttactgtt
     2101 gagcattcat tttaaatgcc agcaaggttt acgcacacgc attctgttac tgcggatggt
     2161 gacctggatt ggtcgctggt cgatgctcga cctgtttgtc atatctttaa ccatgtcgct
     2221 gattaatcgc gatcagatcc tcgcttttac tatgggaccg gctgcgtttt atttcggcgc
     2281 agcggtaatt ttgactattc ttgctgtgga atggctggac agccgcttac tttgggatgc
     2341 acatgagtca ggaaacgccc gcttcgacga ctgaagcgca gattaaaaat aaacgccgta
     2401 tctcaccttt ctggctgctg ccgttcatcg cgctaatgat tgccagttgg ctgatttggg
     2461 acagttatca ggaccggggt aataccgtca ccatcgactt tatgtcggcg gatggtattg
     2521 ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt cggaacagtg caggacatca
     2581 gcctcagcga cgatcttcgt aagattgaag tcaaggtcag catcaagtcc gatatgaaag
     2641 atgcgctgcg cgaagagact cagttctggc tggtgacgcc aaaagcatcg ttggcaggtg
     2701 tctccgggct ggacgccctc gtcggtggga actatatcgg catgatgccc gggaaaggta
     2761 aagagcagga tcactttgtc gcactcgata cccaaccgaa atatcggctg gacaatggcg
     2821 atctgatgat ccacctgcaa gcccccgatc tcggttcgct gaacagcggt tcattggtct
     2881 atttccgcaa gatcccggtg ggaaaagtct acgactatgc catcaatccc aacaagcaag
     2941 gcgtggtgat tgatgtcctg atcgagcggc gttttaccga tctggtgaaa aaaggtagcc
     3001 gtttctggaa cgtttccggc gttgatgcca acgtcagtat cagtggcgcg aaggtgaaac
     3061 tggaaagctt ggcggcactg gttaacggtg cgattgcctt cgattcacca gaagagtcga
     3121 aacctgccga ggcggaagat acctttggtc tgtatgaaga tctggcccac agccagcgtg
     3181 gcgtaataat aaaactggaa ctgccgagtg gggccggatt aaccgccgac tcgacgccgt
     3241 taatgtatca ggggctggaa gtcggacagc tgactaaact ggatttaaat cctggtggta
     3301 aagtcacagg ggaaatgacc gttgatccca gcgtcgttac cctgcttcgt gaaaataccc
     3361 gcatcgaatt acgcaacccg aaattatccc ttagcgatgc taatctcagc gccctgctga
     3421 ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc acgcaaagaa ttcgttgttg
     3481 tgccaggcga aaaagcactg ctgcatgaac ctgatgttct gacgctgacc ctgaccgcgc
     3541 cggaaagtta cggtattgat gcgggtcagc cgctcattct tcacggcgtg caggtaggcc
     3601 aggttatcga tcgtaaactc accagcaaag gcgtcacctt taccgtcgcc atcgagcctc
     3661 agcatcgcga actggtaaaa ggcgatagca aatttgtcgt caacagccgt gtcgatgtga
     3721 aggtggggct ggatggcgtt gagtttctcg gtgccagcgc ctcagaatgg atcaatggcg
     3781 ggatacgtat tctgccgggc gataaaggtg agatgaaagc cagctatcca ctgtatgcca
     3841 atttggaaaa agcgctggag aacagcctta gcgatttacc caccacaacc gtgagtttga
     3901 gtgcagagac gctgccggat gtgcaggcag gatcggtagt gctgtaccgt aaatttgaag
     3961 ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt tgatatcgat ctgcatatta
     4021 agccggagta tcgcaacctt ctgaccagca atagcgtgtt ctgggcagaa ggcggggcga
     4081 aagttcagct gaatggtagt ggtctgaccg tacaggcatc cccgctctcc agagcattaa
     4141 agggagccat tagcttcgac aacctcagcg gtgccagcgc cagtcagcgt aaaggcgata
     4201 agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt tggcgggcag attacgcttc
     4261 acgctttcga tgccggaaaa ctggcggtcg ggatgccaat tcgctatctc ggtattgata
     4321 tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa tgaagtacag gcaaaggcgg
     4381 tactttaccc ggaatatgtc cagacctttg ctcgcggtgg tacgcgcttc tcagtggtca
     4441 caccgcaaat ttcggcagct ggcgttgagc ataatgatac tatcctccag ccgtatatca
     4501 acgtcgaacc aggccggggc aatcctcgcc gcgactttga attacaagag gccaccatta
     4561 ctgattcgcg ttacctggat ggcttaagca ttattgttga agcgccggaa gccggttcgt
     4621 taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt cggtacggtt acaggaatga
     4681 cgctggggac attgtcagat cgcgtgatga ttgcgatgcg catcagtaaa cgctatcaac
     4741 acctggtgcg taacaattcc gtcttctggt tggcatcggg ttacagtctg gactttggtc
     4801 tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt tatccgtggc ggcatcgcct
     4861 tcgccacgcc tccgggtacg ccactggcac cgaaagccca ggaaggcaaa cacttcctgt
     4921 tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac tgcgcttccc aaataatgcc
     4981 cacctcgagc agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     5041 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     5101 tgctgaaagg aggaactata tccggataat tcttgaagac gaaagggcct cgtgatacgc
     5161 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5221 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5281 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5341 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5401 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5461 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5521 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5581 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5641 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5701 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5761 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5821 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5881 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5941 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6001 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6061 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6121 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6181 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6481 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtg
//
