LOCUS       sequence-345243-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB(G707N).
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: pBEL2418
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     256..273
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    288..427
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(353..372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     513..535
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(529..720)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1229..1248
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1229..1247
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1279..1301
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5259..5277)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5273..5320
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5365..5383)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5451..5555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5556..5624,5625..6416)
                     /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(5774..5793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6587..7073,1..102)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
       61 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      181 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      241 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
      301 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
      361 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
      421 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
      481 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
      541 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
      601 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
      661 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
      721 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
      781 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
      841 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
      901 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
      961 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     1021 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     1081 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     1141 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     1201 gctgcccgag atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg
     1261 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat ggctcttaac
     1321 acaccacaaa ttacgccgac aaaaaagata acagtgaggg caatcggcga ggaactgccg
     1381 cgtggtgatt accaacgttg cccgcaatgt gacatgctgt ttagcctgcc cgagataaat
     1441 tctcatcaaa gtgcctattg tccgcgctgt caggcaaaaa ttcgtgacgg gcgcgactgg
     1501 tcgctaacgc gcctggcggc aatggccttc accatgctgt tgcttatgcc gtttgcctgg
     1561 ggcgaaccgc tgttgcatat ctggctgtta ggcattcgta ttgacgccaa cgttatgcaa
     1621 ggcatctggc aaatgaccaa acagggcgat gcgataacgg ggtcgatggt ctttttctgc
     1681 gttattggtg ccccgctcat tctggtgacc tccatagctt atttatggtt tggtaaccga
     1741 ctgggaatga atttacgtcc ggtactgctg atgcttgagc gacttaaaga gtgggtaatg
     1801 ctcgatatct acctggtcgg cattggcgtt gcttctataa aggtacagga ttatgcccat
     1861 atccaggctg gtgtcggctt gttctctttt gtggcgttgg tgattttaac gacggtgacg
     1921 ttgtcacatc ttaatgtcga agaactgtgg gagcgatttt atccgcagcg ccccgctacg
     1981 cgtagggacg agaaacttcg tgtctgtctt gggtgccatt ttaccggcta tccagatcag
     2041 cgtggtcgct gcccgcgttg ccatatcccg ctacgcctgc gtcgccgtca tagtctgcaa
     2101 aaatgctggg cggcgctgtt agcgtcaatc gttttattgt tacctgccaa cctgttgcct
     2161 atttctatca tttatctgaa cggaggacgg caggaagata caattctttc cggaattatg
     2221 tcgctggcaa gtagcaacat tgcggttgca ggaatcgtgt ttatcgccag tattctggta
     2281 ccgtttacta aagtgatcgt catgttcact ttactgttga gcattcattt taaatgccag
     2341 caaggtttac gcacacgcat tctgttactg cggatggtga cctggattgg tcgctggtcg
     2401 atgctcgacc tgtttgtcat atctttaacc atgtcgctga ttaatcgcga tcagatcctc
     2461 gcttttacta tgggaccggc tgcgttttat ttcggcgcag cggtaatttt gactattctt
     2521 gctgtggaat ggctggacag ccgcttactt tgggatgcac atgagtcagg aaacgcccgc
     2581 ttcgacgact gaagcgcaga ttaaaaataa acgccgtatc tcacctttct ggctgctgcc
     2641 gttcatcgcg ctaatgattg ccagttggct gatttgggac agttatcagg accggggtaa
     2701 taccgtcacc atcgacttta tgtcggcgga tggtattgtt ccaggccgta cgcctgttcg
     2761 ttatcagggc gttgaagtcg gaacagtgca ggatatcagc ctcagcgacg atcttcgtaa
     2821 gattgaagtc aaggtcagca tcaagtccga tatgaaagat gcgctgcgcg aagagactca
     2881 gttctggctg gtgacgccaa aagcatcgtt ggcaggtgtc tccgggctgg acgccctcgt
     2941 cggtgggaac tatatcggca tgatgccggg taaaggtaaa gagcaggatc actttgtcgc
     3001 actcgatacc caaccgaaat atcggctgga caatggcgat ctgatgatcc acctgcaagc
     3061 ccccgatctc ggttcgctga acagcggttc attggtctat ttccgcaaga tcccggtggg
     3121 aaaagtctac gactatgcca tcaatcccaa caagcaaggc gtggtgattg atgtcctgat
     3181 cgagcggcgt tttaccgatc tggtgaaaaa aggtagccgt ttctggaacg tttccggcgt
     3241 tgatgccaac gtcagtatca gtggcgcgaa ggtgaaactg gaaagcctgg cggcactggt
     3301 taacggtgcg attgccttcg attcaccaga agagtcgaaa cctgccgagg cggaagatac
     3361 ctttggtctg tatgaagatc tggcccacag ccagcgtggc gtaataataa aactggaact
     3421 gccgagtggg gccggattaa ccgccgactc gacgccgtta atgtatcagg ggctggaagt
     3481 cggacagctg actaaactgg atttaaatcc tggtggtaaa gtcacagggg aaatgaccgt
     3541 tgatcccagc gtcgttaccc tgcttcgtga aaatacccgc atcgaattac gcaacccgaa
     3601 attatccctt agcgatgcta atctcagcgc cctgctgacc ggcaaaacct tcgagctggt
     3661 gcccggcgat ggcgagccac gcaaagagtt cgttgttgtg ccaggcgaaa aagcactgct
     3721 gcatgaacct gatgttctga cgctgaccct gaccgcgccg gaaagttacg gtattgatgc
     3781 gggtcagccg ctcattcttc acggcgtgca ggtaggccag gttatcgatc gtaaactcac
     3841 cagcaaaggc gtcaccttta ccgtcgccat cgagcctcag catcgcgaac tggtaaaagg
     3901 cgatagcaaa tttgtcgtca acagccgtgt cgatgtgaag gtggggctgg atggcgttga
     3961 gtttctcggt gccagcgcct cagaatggat caatggcggg atacgtattc tgccgggcga
     4021 taaaggtgag atgaaagcca gctatccact gtatgccaat ttggaaaaag cgctggagaa
     4081 cagccttagc gatttaccca ccacaaccgt gagtttgagt gcagagacgc tgccggatgt
     4141 gcaggcagga tcggtagtgc tgtaccgtaa atttgaagtt ggtgaagtga ttaccgtgcg
     4201 tccgcgagct aacgcgtttg atatcgatct gcatattaag ccggagtatc gcaaccttct
     4261 gaccagcaat agcgtgttct gggcagaagg cggggcgaaa gttcagctga atggtagtgg
     4321 tctgaccgta caggcatccc cgctctccag agcattaaag ggagccatta gcttcgacaa
     4381 cctcagcggt gccagcgcca gtcagcgtaa aggcgataag cgtattctgt atgcttccga
     4441 aacagcggcc cgtgcggttg gcgggcagat tacgcttcac gctttcgatg ccggaaaact
     4501 ggcggtcggg atgccaattc gctatctcgg tattgatatc gggcaaatcc agacgctgga
     4561 tctgattacc gcacgcaatg aagtacaggc aaaggcggta ctttacccgg aatatgtcca
     4621 gacctttgct cgcggtggta cgcgcttctc agtggtcaca ccgcaaattt cggcagctaa
     4681 tgttgagcat cttgatacta tcctccagcc gtatatcaac gtcgaaccag gccggggcaa
     4741 tcctcgccgc gactttgaat tacaagaggc caccattact gattcgcgtt acctggatgg
     4801 cttaagcatt attgttgaag cgccggaagc cggttcgtta ggcatcggta cgcctgtgct
     4861 gttccgtggt ctggaagtcg gtacggttac aggaatgacg ctggggacat tgtcagatcg
     4921 cgtgatgatt gcgatgcgca tcagtaaacg ctatcaacac ctggtgcgta acaattccgt
     4981 cttctggttg gcatcgggtt acagtctgga ctttggtctg acgggcggcg tagtgaaaac
     5041 cggcaccttt aaccagttta tccgtggcgg catcgccttc gccacgcctc cgggtacgcc
     5101 actggcaccg aaagcccagg aaggcaaaca cttcctgttg caggaaagtg aaccgaaaga
     5161 gtggcgtgaa tggggaactg cgcttcccaa ataatgccca cctcgagcag atccggctgc
     5221 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     5281 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     5341 cggataattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5401 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5461 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5521 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5581 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5641 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5701 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5761 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     5821 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5881 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5941 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6001 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6061 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     6121 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6181 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6241 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6301 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6361 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6421 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6481 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6721 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt cca
//
