LOCUS       sequence-337621-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant A473N.
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: pBEL1863
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..88)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    174..313
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     229..248
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(328..345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(499..1087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(579..598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1258..2049,2050..2118))
                     /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     1881..1900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2119..2223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2291..2309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2354..2401)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2397..2415
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             6373..6395
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6426..6445)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6427..6445)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(6954..7073,1..72)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt
       61 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
      121 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
      181 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
      241 agattgtact gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa
      301 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      361 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      421 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      481 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      541 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      601 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      661 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      721 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      781 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      841 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      901 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
      961 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1021 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1081 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1141 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1201 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1261 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1321 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1381 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1441 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1501 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1561 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1621 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1681 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1741 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1801 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1861 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1921 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1981 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2041 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2101 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2161 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2221 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2281 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattat ccggatatag
     2341 ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     2401 gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg
     2461 gatctgctcg aggtgggcat tatttgggaa gcgcagttcc ccattcacgc cactctttcg
     2521 gttcactttc ctgcaacagg aagtgtttgc cttcctgggc tttcggtgcc agtggcgtac
     2581 ccggaggcgt ggcgaaggcg atgccgccac ggataaactg gttaaaggtg ccggttttca
     2641 ctacgccgcc cgtcagacca aagtccagac tgtaacccga tgccaaccag aagacggaat
     2701 tgttacgcac caggtgttga tagcgtttac tgatgcgcat cgcaatcatc acgcgatctg
     2761 acaatgtccc cagcgtcatt cctgtaaccg taccgacttc cagaccacgg aacagcacag
     2821 gcgtaccgat gcctaacgaa ccggcttccg gcgcttcaac aataatgctt aagccatcca
     2881 ggtaccgcga atcagtaatg gtggcctctt gtaattcaaa gtcgcggcga ggattgcccc
     2941 ggcctggttc gacgttgata tacggctgga ggatagtatc aagatgctca acgccagctg
     3001 ccgaaatttg cggtgtgacc actgagaagc gcgtaccacc gcgagcaaag gtctggacat
     3061 attccgggta aagtaccgcc tttgcctgta cttcattgcg tgcggtaatc agatccagcg
     3121 tctggatttg cccgatgtca ataccgagat agcgaattgg catcccgacc gccagttttc
     3181 cggcatcgaa agcgtgaagc gtaatctgcc cgccaaccgc acgggccgct gtttcggaag
     3241 catacagaat acgcttatcg cctttacgct gactggcgct ggcaccgctg aggttgtcga
     3301 agctaatggc tccctttaga gctctggaga gcggggatgc ctgtacggtc agaccactac
     3361 cattcagctg aactttcgcc ccgccttctg cccagaacac gctattgctg gtcagaaggt
     3421 tgcgatactc cggcttaata tgcagatcga tgtcaaacgc gttagctcgc ggacgcacgg
     3481 taatcacttc accaacttca aatttacggt acagcactac cgatcctgcc tgcacatccg
     3541 gcagcgtctc tgcactcaaa ctcacggttg tggtgggtaa atcgctaagg ctgttctcca
     3601 gcgctttttc caaattggca tacagtggat agctggcttt catctcacct ttatcgcccg
     3661 gcagaatacg tatcccgcca ttgatccatt ctgaattgct ggcaccgaga aactcaacgc
     3721 catccagccc caccttcaca tcgacacggc tgttgacgac aaatttgcta tcgcctttta
     3781 ccagttcgcg atgctgaggc tcgatggcga cggtaaaggt gacgcctttg ctggtgagtt
     3841 tacgatcgat aacctggcct acctgcacgc cgtgaagaat gagcggctga cccgcatcaa
     3901 taccgtaact ttccggcgcg gtcagggtca gcgtcagaac atcaggttca tgcagcagtg
     3961 ctttttcgcc tggcacaaca acgaactctt tgcgtggctc gccatcgccg ggcaccagct
     4021 cgaaggtttt gccggtcagc agggcgctga gattagcatc gctaagggat aatttcgggt
     4081 tgcgtaattc gatgcgggta ttttcacgaa gcagggtaac gacgctggga tcaacggtca
     4141 tttcccctgt gactttacca ccaggattta aatccagttt agtcagctgt ccgacttcca
     4201 gcccctgata cattaacggc gtcgagtcgg cggttaatcc ggccccactc ggcagttcca
     4261 gttttattat tacgccacgc tggctgtggg ccagatcttc atacagacca aaggtatctt
     4321 ccgcctcggc aggtttcgac tcttctggtg aatcgaaggc aatcgcaccg ttaaccagtg
     4381 ccgccaggct ttccagtttc accttcgcgc cactgatact gacgttggca tcaacgccgg
     4441 aaacgttcca gaaacggcta ccttttttca ccagatcggt aaaacgccgc tcgatcagga
     4501 catcaatcac cacgccttgc ttgttgggat tgatggcata gtcgtagact tttcccaccg
     4561 ggatcttgcg gaaatagacc aatgaaccgc tgttcagcga accgagatcg ggggcttgca
     4621 ggtggatcat cagatcgcca ttgtccagcc gatatttcgg ttgggtatcg agtgcgacaa
     4681 agtgatcctg ctctttacct ttacccggca tcatgccgat atagttccca ccgacgaggg
     4741 cgtccagccc ggagacacct gccaacgatg cttttggcgt caccagccag aactgagtct
     4801 cttcgcgcag cgcatctttc atatcggact tgatgctgac cttgacttca atcttacgaa
     4861 gatcgtcgct gaggctgatg tcctgcactg ttccgacttc aacgccctga taacgaacag
     4921 gcgtacggcc tggaacaata ccatccgccg acataaagtc gatggtgacg gtattacccc
     4981 ggtcctgata actgtcccaa atcagccaac tggcaatcat tagcgcgatg aacggcagca
     5041 gccagaaagg tgagatacgg cgtttatttt taatctgcgc ttcagtcgtc gaagcgggcg
     5101 tttcctgact catgtgcatc ccaaagtaag cggctgtcca gccattccac agcaagaata
     5161 gtcaaaatta ccgctgcgcc gaaataaaac gcagccggtc ccatagtaaa agcgaggatc
     5221 tgatcgcgat taatcagcga catggttaaa gatatgacaa acaggtcgag catcgaccag
     5281 cgaccaatcc aggtcaccat ccgcagtaac agaatgcgtg tgcgtaaacc ttgctggcat
     5341 ttaaaatgaa tgctcaacag taaagtgaac atgacgatca ctttagtaaa cgggaccaga
     5401 atactggcga taaacacgat tcctgcaacc gcaatgttgc tacttgccag cgacataatt
     5461 ccggaaagaa ttgtatcttc ctgccgtcct ccgttcagat aaatgataga aataggcaac
     5521 aggttggcag gtaacaataa aacgattgac gctaacagcg ccgcccagca tttttgcaga
     5581 ctatgacggc gacgcaggcg tagcgggata tggcaacgcg ggcagcgacc acgctgatct
     5641 ggatagccgg taaaatggca ccctaggcag acacgaagtt tctcgtccct acgcgtagcg
     5701 gggcgctgcg gataaaatcg ctcccacagt tcttcgacat taagatgtga caacgtcacc
     5761 gtcgttaaaa tcaccaacgc cacaaaagag aacaagccga caccagcctg gatatgggca
     5821 taatcctgta cctttataga agcaacgcca atgccgacca ggtagatatc gagcattacc
     5881 cactctttaa gtcgctcaag catcagcagt accggacgta aattcattcc cagtcggtta
     5941 ccaaaccata aataagctat ggaggtcacc agaatgagcg gggcaccaat aacgcagaaa
     6001 aagaccatcg accccgttat cgcatcgccc tgtttggtca tttgccagat gccttgcata
     6061 acgttggcgt caatacgaat gcctaacagc cagatatgca acagcggttc gccccaggca
     6121 aacggcataa gcaacagcat ggtgaaggcc attgccgcca ggcgcgttag cgaccagtcg
     6181 cgcccgtcac gaatttttgc ctgacagcgc ggacaatagg cactttgatg agaatttatc
     6241 tcgggcaggc taaacagcat gtcacattgc gggcaacgtt ggtaatcacc acgcggcagt
     6301 tcctcgccga ttgccctcac tgttatcttt tttgtcggcg taatttgtgg tgtgttaaga
     6361 gccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg aaaccgttgt
     6421 ggtctcccta tagtgagtcg tattaatttc gcgggatcga gatctcgggc agcgttgggt
     6481 cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     6541 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     6601 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     6661 aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag
     6721 gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac
     6781 cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac
     6841 gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt
     6901 tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca
     6961 aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc
     7021 tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa tcg
//
