LOCUS       sequence-334344-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant I834N.
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: pBEL1874
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     242..259
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    274..413
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(339..358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     499..521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(515..706)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1215..1234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1215..1233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1265..1287
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5245..5263)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5259..5306
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5351..5369)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5437..5541
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5542..5610,5611..6402)
                     /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(5760..5779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6573..7073,1..88)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(7062..7073,1..8)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
       61 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      121 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      181 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      241 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
      301 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
      361 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
      421 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
      481 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
      541 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
      601 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
      661 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
      721 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
      781 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
      841 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
      901 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
      961 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     1021 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     1081 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     1141 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct
     1201 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
     1261 taattttgtt taactttaag aaggagatat acatatggct cttaacacac cacaaattac
     1321 gccgacaaaa aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca
     1381 acgttgcccg caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc
     1441 ctattgtccg cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct
     1501 ggcggcaatg gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt
     1561 gcatatctgg ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat
     1621 gaccaaacag ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc
     1681 gctcattctg gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt
     1741 acgtccggta ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct
     1801 ggtcggcatt ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt
     1861 cggcttgttc tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa
     1921 tgtcgaagaa ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa
     1981 acttcgtgtc tgcctagggt gccattttac cggctatcca gatcagcgtg gtcgctgccc
     2041 gcgttgccat atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc
     2101 gctgttagcg tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta
     2161 tctgaacgga ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag
     2221 caacattgcg gttgcaggaa tcgtgtttat cgccagtatt ctggtcccgt ttactaaagt
     2281 gatcgtcatg ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac
     2341 acgcattctg ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt
     2401 tgtcatatct ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg
     2461 accggctgcg ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct
     2521 ggacagccgc ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag
     2581 cgcagattaa aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa
     2641 tgattgccag ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg
     2701 actttatgtc ggcggatggt attgttccag gccgtacgcc tgttcgttat cagggcgttg
     2761 aagtcggaac agtgcaggac atcagcctca gcgacgatct tcgtaagatt gaagtcaagg
     2821 tcagcatcaa gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga
     2881 cgccaaaagc atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata
     2941 tcggcatgat gcccgggaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac
     3001 cgaaatatcg gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt
     3061 cgctgaacag cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact
     3121 atgccatcaa tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta
     3181 ccgatctggt gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca
     3241 gtatcagtgg cgcgaaggtg aaactggaaa gcttggcggc actggttaac ggtgcgattg
     3301 ccttcgattc accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg
     3361 aagatctggc ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg
     3421 gattaaccgc cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta
     3481 aactggattt aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg
     3541 ttaccctgct tcgtgaaaat acccgcatcg aattacgcaa cccgaaatta tcccttagcg
     3601 atgctaatct cagcgccctg ctgaccggca aaaccttcga gctggtgccc ggcgatggcg
     3661 agccacgcaa agaattcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg
     3721 ttctgacgct gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca
     3781 ttcttcacgg cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca
     3841 cctttaccgt cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg
     3901 tcgtcaacag ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca
     3961 gcgcctcaga atggatcaat ggcgggatcc gtattctgcc gggcgataaa ggtgagatga
     4021 aagccagcta tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt
     4081 tacccaccac aaccgtgagt ttgagtgcag agacgctgcc ggatgtgcag gcaggatcgg
     4141 tagtgctgta ccgtaaattt gaagttggtg aagtgattac cgtgcgtccg cgagctaacg
     4201 cgtttgacat cgatctgcat attaagccgg agtatcgcaa ccttctgacc agcaatagcg
     4261 tgttctgggc agaaggcggg gcgaaagttc agctgaatgg tagtggtctg accgtacagg
     4321 catccccgct ctccagagct ctaaagggag ccattagctt cgacaacctc agcggtgcca
     4381 gcgccagtca gcgtaaaggc gataagcgta ttctgtatgc ttccgaaaca gcggcccgtg
     4441 cggttggcgg gcagattacg cttcacgctt tcgatgccgg aaaactggcg gtcgggatgc
     4501 caattcgcta tctcggtatt gacatcgggc aaatccagac gctggatctg attaccgcac
     4561 gcaatgaagt acaggcaaag gcggtacttt acccggaata tgtccagacc tttgctcgcg
     4621 gtggtacgcg cttctcagtg gtcacaccgc aaatttcggc agctggcgtt gagcatcttg
     4681 atactatcct ccagccgtat atcaacgtcg aaccaggccg gggcaatcct cgccgcgact
     4741 ttgaattaca agaggccacc attactgatt cgcggtacct ggatggctta agcattattg
     4801 ttgaagcgcc ggaagccggt tcgttaggca tcggtacgcc tgtgctgttc cgtggtctgg
     4861 aagtcggtac ggttacagga atgacgctgg ggacattgtc agatcgcgtg atgattgcga
     4921 tgcgcatcag taaacgctat caacacctgg tgcgtaacaa ttccgtcttc tggttggcat
     4981 cgggttacag tctggacttt ggtctgacgg gcggcgtagt gaaaaccggc acctttaacc
     5041 agtttaatcg tggcggcatc gccttcgcca cgcctccggg tacgccactg gcaccgaaag
     5101 cccaggaagg caaacacttc ctgttgcagg aaagtgaacc gaaagagtgg cgtgaatggg
     5161 gaactgcgct tcccaaataa tgcccacctc gagcagatcc ggctgctaac aaagcccgaa
     5221 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     5281 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taattcttga
     5341 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5401 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5461 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5521 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5581 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5641 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5701 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5761 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     5821 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5881 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5941 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6001 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6061 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     6121 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6181 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6241 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6301 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6361 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6421 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6481 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6541 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6601 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6661 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6721 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6781 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6841 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6901 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6961 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7021 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctg
//
