LOCUS       sequence-334332-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant I714N.
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: pBEL1870
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    158..297
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(223..242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     383..405
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(399..590)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1099..1118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1099..1117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1149..1171
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5129..5147)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5143..5190
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5235..5253)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5321..5425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5426..5494,5495..6286)
                     /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(5644..5663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6457..7045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6946..6965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
       61 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      121 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
      181 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
      241 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
      301 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      361 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      421 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
      481 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
      541 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
      601 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
      661 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
      721 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
      781 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
      841 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
      901 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
      961 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     1021 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag
     1081 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
     1141 gaaataattt tgtttaactt taagaaggag atatacatat ggctcttaac acaccacaaa
     1201 ttacgccgac aaaaaagata acagtgaggg caatcggcga ggaactgccg cgtggtgatt
     1261 accaacgttg cccgcaatgt gacatgctgt ttagcctgcc cgagataaat tctcatcaaa
     1321 gtgcctattg tccgcgctgt caggcaaaaa ttcgtgacgg gcgcgactgg tcgctaacgc
     1381 gcctggcggc aatggccttc accatgctgt tgcttatgcc gtttgcctgg ggcgaaccgc
     1441 tgttgcatat ctggctgtta ggcattcgta ttgacgccaa cgttatgcaa ggcatctggc
     1501 aaatgaccaa acagggcgat gcgataacgg ggtcgatggt ctttttctgc gttattggtg
     1561 ccccgctcat tctggtgacc tccatagctt atttatggtt tggtaaccga ctgggaatga
     1621 atttacgtcc ggtactgctg atgcttgagc gacttaaaga gtgggtaatg ctcgatatct
     1681 acctggtcgg cattggcgtt gcttctataa aggtacagga ttatgcccat atccaggctg
     1741 gtgtcggctt gttctctttt gtggcgttgg tgattttaac gacggtgacg ttgtcacatc
     1801 ttaatgtcga agaactgtgg gagcgatttt atccgcagcg ccccgctacg cgtagggacg
     1861 agaaacttcg tgtctgccta gggtgccatt ttaccggcta tccagatcag cgtggtcgct
     1921 gcccgcgttg ccatatcccg ctacgcctgc gtcgccgtca tagtctgcaa aaatgctggg
     1981 cggcgctgtt agcgtcaatc gttttattgt tacctgccaa cctgttgcct atttctatca
     2041 tttatctgaa cggaggacgg caggaagata caattctttc cggaattatg tcgctggcaa
     2101 gtagcaacat tgcggttgca ggaatcgtgt ttatcgccag tattctggtc ccgtttacta
     2161 aagtgatcgt catgttcact ttactgttga gcattcattt taaatgccag caaggtttac
     2221 gcacacgcat tctgttactg cggatggtga cctggattgg tcgctggtcg atgctcgacc
     2281 tgtttgtcat atctttaacc atgtcgctga ttaatcgcga tcagatcctc gcttttacta
     2341 tgggaccggc tgcgttttat ttcggcgcag cggtaatttt gactattctt gctgtggaat
     2401 ggctggacag ccgcttactt tgggatgcac atgagtcagg aaacgcccgc ttcgacgact
     2461 gaagcgcaga ttaaaaataa acgccgtatc tcacctttct ggctgctgcc gttcatcgcg
     2521 ctaatgattg ccagttggct gatttgggac agttatcagg accggggtaa taccgtcacc
     2581 atcgacttta tgtcggcgga tggtattgtt ccaggccgta cgcctgttcg ttatcagggc
     2641 gttgaagtcg gaacagtgca ggacatcagc ctcagcgacg atcttcgtaa gattgaagtc
     2701 aaggtcagca tcaagtccga tatgaaagat gcgctgcgcg aagagactca gttctggctg
     2761 gtgacgccaa aagcatcgtt ggcaggtgtc tccgggctgg acgccctcgt cggtgggaac
     2821 tatatcggca tgatgcccgg gaaaggtaaa gagcaggatc actttgtcgc actcgatacc
     2881 caaccgaaat atcggctgga caatggcgat ctgatgatcc acctgcaagc ccccgatctc
     2941 ggttcgctga acagcggttc attggtctat ttccgcaaga tcccggtggg aaaagtctac
     3001 gactatgcca tcaatcccaa caagcaaggc gtggtgattg atgtcctgat cgagcggcgt
     3061 tttaccgatc tggtgaaaaa aggtagccgt ttctggaacg tttccggcgt tgatgccaac
     3121 gtcagtatca gtggcgcgaa ggtgaaactg gaaagcttgg cggcactggt taacggtgcg
     3181 attgccttcg attcaccaga agagtcgaaa cctgccgagg cggaagatac ctttggtctg
     3241 tatgaagatc tggcccacag ccagcgtggc gtaataataa aactggaact gccgagtggg
     3301 gccggattaa ccgccgactc gacgccgtta atgtatcagg ggctggaagt cggacagctg
     3361 actaaactgg atttaaatcc tggtggtaaa gtcacagggg aaatgaccgt tgatcccagc
     3421 gtcgttaccc tgcttcgtga aaatacccgc atcgaattac gcaacccgaa attatccctt
     3481 agcgatgcta atctcagcgc cctgctgacc ggcaaaacct tcgagctggt gcccggcgat
     3541 ggcgagccac gcaaagaatt cgttgttgtg ccaggcgaaa aagcactgct gcatgaacct
     3601 gatgttctga cgctgaccct gaccgcgccg gaaagttacg gtattgatgc gggtcagccg
     3661 ctcattcttc acggcgtgca ggtaggccag gttatcgatc gtaaactcac cagcaaaggc
     3721 gtcaccttta ccgtcgccat cgagcctcag catcgcgaac tggtaaaagg cgatagcaaa
     3781 tttgtcgtca acagccgtgt cgatgtgaag gtggggctgg atggcgttga gtttctcggt
     3841 gccagcgcct cagaatggat caatggcggg atacgtattc tgccgggcga taaaggtgag
     3901 atgaaagcca gctatccact gtatgccaat ttggaaaaag cgctggagaa cagccttagc
     3961 gatttaccca ccacaaccgt gagtttgagt gcagagacgc tgccggatgt gcaggcagga
     4021 tcggtagtgc tgtaccgtaa atttgaagtt ggtgaagtga ttaccgtgcg tccgcgagct
     4081 aacgcgtttg atatcgatct gcatattaag ccggagtatc gcaaccttct gaccagcaat
     4141 agcgtgttct gggcagaagg cggggcgaaa gttcagctga atggtagtgg tctgaccgta
     4201 caggcatccc cgctctccag agcattaaag ggagccatta gcttcgacaa cctcagcggt
     4261 gccagcgcca gtcagcgtaa aggcgataag cgtattctgt atgcttccga aacagcggcc
     4321 cgtgcggttg gcgggcagat tacgcttcac gctttcgatg ccggaaaact ggcggtcggg
     4381 atgccaattc gctatctcgg tattgatatc gggcaaatcc agacgctgga tctgattacc
     4441 gcacgcaatg aagtacaggc aaaggcggta ctttacccgg aatatgtcca gacctttgct
     4501 cgcggtggta cgcgcttctc agtggtcaca ccgcaaattt cggcagctgg cgttgagcat
     4561 cttgatacta acctccagcc gtatatcaac gtcgaaccag gccggggcaa tcctcgccgc
     4621 gactttgaat tacaagaggc caccattact gattcgcgtt acctggatgg cttaagcatt
     4681 attgttgaag cgccggaagc cggttcgtta ggcatcggta cgcctgtgct gttccgtggt
     4741 ctggaagtcg gtacggttac aggaatgacg ctggggacat tgtcagatcg cgtgatgatt
     4801 gcgatgcgca tcagtaaacg ctatcaacac ctggtgcgta acaattccgt cttctggttg
     4861 gcatcgggtt acagtctgga ctttggtctg acgggcggcg tagtgaaaac cggcaccttt
     4921 aaccagttta tccgtggcgg catcgccttc gccacgcctc cgggtacgcc actggcaccg
     4981 aaagcccagg aaggcaaaca cttcctgttg caggaaagtg aaccgaaaga gtggcgtgaa
     5041 tggggaactg cgcttcccaa ataatgccca cctcgagcag atccggctgc taacaaagcc
     5101 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     5161 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggataattc
     5221 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5281 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5341 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5401 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5461 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5521 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5581 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5641 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     5701 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5761 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5821 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5881 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5941 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     6001 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6061 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6121 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6181 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6241 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6301 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6601 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggt
//
