LOCUS       sequence-334365-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant L715N.
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: pBEL1871
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     178..195
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    210..349
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(275..294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     435..457
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(451..642)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1151..1170
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1151..1169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1201..1223
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5181..5199)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5195..5242
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5287..5305)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5373..5477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5478..5546,5547..6338)
                     /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(5696..5715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6509..7073,1..24)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6998..7017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
       61 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      121 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      181 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
      241 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      301 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
      361 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      421 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      481 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
      541 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
      601 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
      661 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
      721 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
      781 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
      841 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
      901 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
      961 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     1021 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     1081 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatctcgat
     1141 cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc tagaaataat
     1201 tttgtttaac tttaagaagg agatatacat atggctctta acacaccaca aattacgccg
     1261 acaaaaaaga taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt
     1321 tgcccgcaat gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat
     1381 tgtccgcgct gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg
     1441 gcaatggcct tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat
     1501 atctggctgt taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc
     1561 aaacagggcg atgcgataac ggggtcgatg gtctttttct gcgttattgg tgccccgctc
     1621 attctggtga cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt
     1681 ccggtactgc tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc
     1741 ggcattggcg ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc
     1801 ttgttctctt ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc
     1861 gaagaactgt gggagcgatt ttatccgcag cgccccgcta cgcgtaggga cgagaaactt
     1921 cgtgtctgcc tagggtgcca ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt
     1981 tgccatatcc cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg
     2041 ttagcgtcaa tcgttttatt gttacctgcc aacctgttgc ctatttctat catttatctg
     2101 aacggaggac ggcaggaaga tacaattctt tccggaatta tgtcgctggc aagtagcaac
     2161 attgcggttg caggaatcgt gtttatcgcc agtattctgg tcccgtttac taaagtgatc
     2221 gtcatgttca ctttactgtt gagcattcat tttaaatgcc agcaaggttt acgcacacgc
     2281 attctgttac tgcggatggt gacctggatt ggtcgctggt cgatgctcga cctgtttgtc
     2341 atatctttaa ccatgtcgct gattaatcgc gatcagatcc tcgcttttac tatgggaccg
     2401 gctgcgtttt atttcggcgc agcggtaatt ttgactattc ttgctgtgga atggctggac
     2461 agccgcttac tttgggatgc acatgagtca ggaaacgccc gcttcgacga ctgaagcgca
     2521 gattaaaaat aaacgccgta tctcaccttt ctggctgctg ccgttcatcg cgctaatgat
     2581 tgccagttgg ctgatttggg acagttatca ggaccggggt aataccgtca ccatcgactt
     2641 tatgtcggcg gatggtattg ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt
     2701 cggaacagtg caggacatca gcctcagcga cgatcttcgt aagattgaag tcaaggtcag
     2761 catcaagtcc gatatgaaag atgcgctgcg cgaagagact cagttctggc tggtgacgcc
     2821 aaaagcatcg ttggcaggtg tctccgggct ggacgccctc gtcggtggga actatatcgg
     2881 catgatgccc gggaaaggta aagagcagga tcactttgtc gcactcgata cccaaccgaa
     2941 atatcggctg gacaatggcg atctgatgat ccacctgcaa gcccccgatc tcggttcgct
     3001 gaacagcggt tcattggtct atttccgcaa gatcccggtg ggaaaagtct acgactatgc
     3061 catcaatccc aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc gttttaccga
     3121 tctggtgaaa aaaggtagcc gtttctggaa cgtttccggc gttgatgcca acgtcagtat
     3181 cagtggcgcg aaggtgaaac tggaaagctt ggcggcactg gttaacggtg cgattgcctt
     3241 cgattcacca gaagagtcga aacctgccga ggcggaagat acctttggtc tgtatgaaga
     3301 tctggcccac agccagcgtg gcgtaataat aaaactggaa ctgccgagtg gggccggatt
     3361 aaccgccgac tcgacgccgt taatgtatca ggggctggaa gtcggacagc tgactaaact
     3421 ggatttaaat cctggtggta aagtcacagg ggaaatgacc gttgatccca gcgtcgttac
     3481 cctgcttcgt gaaaataccc gcatcgaatt acgcaacccg aaattatccc ttagcgatgc
     3541 taatctcagc gccctgctga ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc
     3601 acgcaaagaa ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac ctgatgttct
     3661 gacgctgacc ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc cgctcattct
     3721 tcacggcgtg caggtaggcc aggttatcga tcgtaaactc accagcaaag gcgtcacctt
     3781 taccgtcgcc atcgagcctc agcatcgcga actggtaaaa ggcgatagca aatttgtcgt
     3841 caacagccgt gtcgatgtga aggtggggct ggatggcgtt gagtttctcg gtgccagcgc
     3901 ctcagaatgg atcaatggcg ggatacgtat tctgccgggc gataaaggtg agatgaaagc
     3961 cagctatcca ctgtatgcca atttggaaaa agcgctggag aacagcctta gcgatttacc
     4021 caccacaacc gtgagtttga gtgcagagac gctgccggat gtgcaggcag gatcggtagt
     4081 gctgtaccgt aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt
     4141 tgatatcgat ctgcatatta agccggagta tcgcaacctt ctgaccagca atagcgtgtt
     4201 ctgggcagaa ggcggggcga aagttcagct gaatggtagt ggtctgaccg tacaggcatc
     4261 cccgctctcc agagcattaa agggagccat tagcttcgac aacctcagcg gtgccagcgc
     4321 cagtcagcgt aaaggcgata agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt
     4381 tggcgggcag attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat
     4441 tcgctatctc ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa
     4501 tgaagtacag gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg
     4561 tacgcgcttc tcagtggtca caccgcaaat ttcggcagct ggcgttgagc atcttgatac
     4621 tatcaatcag ccgtatatca acgtcgaacc aggccggggc aatcctcgcc gcgactttga
     4681 attacaagag gccaccatta ctgattcgcg ttacctggat ggcttaagca ttattgttga
     4741 agcgccggaa gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt
     4801 cggtacggtt acaggaatga cgctggggac attgtcagat cgcgtgatga ttgcgatgcg
     4861 catcagtaaa cgctatcaac acctggtgcg taacaattcc gtcttctggt tggcatcggg
     4921 ttacagtctg gactttggtc tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt
     4981 tatccgtggc ggcatcgcct tcgccacgcc tccgggtacg ccactggcac cgaaagccca
     5041 ggaaggcaaa cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac
     5101 tgcgcttccc aaataatgcc cacctcgagc agatccggct gctaacaaag cccgaaagga
     5161 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     5221 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac
     5281 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5341 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5401 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5461 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5521 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5581 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5641 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5701 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5761 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5821 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5881 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5941 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6001 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6061 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6121 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6181 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6241 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6301 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6361 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6421 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6481 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6541 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6601 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6661 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6721 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6781 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6841 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6901 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6961 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7021 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcg
//
