LOCUS       sequence-345253-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB(A705N).
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: pBEL2416
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     289..308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     542..559
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    574..713
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(639..658)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     799..821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(815..1006)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1515..1534
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1515..1533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1565..1587
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5545..5563)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5559..5606
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5651..5669)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5737..5841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5842..5910,5911..6702)
                     /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(6060..6079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6873..7073,1..388)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
       61 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      421 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      481 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      541 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
      601 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
      661 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
      721 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
      781 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
      841 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
      901 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
      961 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     1021 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     1081 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     1141 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     1201 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     1261 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     1321 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     1381 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     1441 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct
     1501 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
     1561 taattttgtt taactttaag aaggagatat acatatggct cttaacacac cacaaattac
     1621 gccgacaaaa aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca
     1681 acgttgcccg caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc
     1741 ctattgtccg cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct
     1801 ggcggcaatg gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt
     1861 gcatatctgg ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat
     1921 gaccaaacag ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc
     1981 gctcattctg gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt
     2041 acgtccggta ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct
     2101 ggtcggcatt ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt
     2161 cggcttgttc tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa
     2221 tgtcgaagaa ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa
     2281 acttcgtgtc tgtcttgggt gccattttac cggctatcca gatcagcgtg gtcgctgccc
     2341 gcgttgccat atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc
     2401 gctgttagcg tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta
     2461 tctgaacgga ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag
     2521 caacattgcg gttgcaggaa tcgtgtttat cgccagtatt ctggtaccgt ttactaaagt
     2581 gatcgtcatg ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac
     2641 acgcattctg ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt
     2701 tgtcatatct ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg
     2761 accggctgcg ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct
     2821 ggacagccgc ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag
     2881 cgcagattaa aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa
     2941 tgattgccag ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg
     3001 actttatgtc ggcggatggt attgttccag gccgtacgcc tgttcgttat cagggcgttg
     3061 aagtcggaac agtgcaggat atcagcctca gcgacgatct tcgtaagatt gaagtcaagg
     3121 tcagcatcaa gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga
     3181 cgccaaaagc atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata
     3241 tcggcatgat gccgggtaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac
     3301 cgaaatatcg gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt
     3361 cgctgaacag cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact
     3421 atgccatcaa tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta
     3481 ccgatctggt gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca
     3541 gtatcagtgg cgcgaaggtg aaactggaaa gcctggcggc actggttaac ggtgcgattg
     3601 ccttcgattc accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg
     3661 aagatctggc ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg
     3721 gattaaccgc cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta
     3781 aactggattt aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg
     3841 ttaccctgct tcgtgaaaat acccgcatcg aattacgcaa cccgaaatta tcccttagcg
     3901 atgctaatct cagcgccctg ctgaccggca aaaccttcga gctggtgccc ggcgatggcg
     3961 agccacgcaa agagttcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg
     4021 ttctgacgct gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca
     4081 ttcttcacgg cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca
     4141 cctttaccgt cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg
     4201 tcgtcaacag ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca
     4261 gcgcctcaga atggatcaat ggcgggatac gtattctgcc gggcgataaa ggtgagatga
     4321 aagccagcta tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt
     4381 tacccaccac aaccgtgagt ttgagtgcag agacgctgcc ggatgtgcag gcaggatcgg
     4441 tagtgctgta ccgtaaattt gaagttggtg aagtgattac cgtgcgtccg cgagctaacg
     4501 cgtttgatat cgatctgcat attaagccgg agtatcgcaa ccttctgacc agcaatagcg
     4561 tgttctgggc agaaggcggg gcgaaagttc agctgaatgg tagtggtctg accgtacagg
     4621 catccccgct ctccagagca ttaaagggag ccattagctt cgacaacctc agcggtgcca
     4681 gcgccagtca gcgtaaaggc gataagcgta ttctgtatgc ttccgaaaca gcggcccgtg
     4741 cggttggcgg gcagattacg cttcacgctt tcgatgccgg aaaactggcg gtcgggatgc
     4801 caattcgcta tctcggtatt gatatcgggc aaatccagac gctggatctg attaccgcac
     4861 gcaatgaagt acaggcaaag gcggtacttt acccggaata tgtccagacc tttgctcgcg
     4921 gtggtacgcg cttctcagtg gtcacaccgc aaatttcgaa tgctggcgtt gagcatcttg
     4981 atactatcct ccagccgtat atcaacgtcg aaccaggccg gggcaatcct cgccgcgact
     5041 ttgaattaca agaggccacc attactgatt cgcgttacct ggatggctta agcattattg
     5101 ttgaagcgcc ggaagccggt tcgttaggca tcggtacgcc tgtgctgttc cgtggtctgg
     5161 aagtcggtac ggttacagga atgacgctgg ggacattgtc agatcgcgtg atgattgcga
     5221 tgcgcatcag taaacgctat caacacctgg tgcgtaacaa ttccgtcttc tggttggcat
     5281 cgggttacag tctggacttt ggtctgacgg gcggcgtagt gaaaaccggc acctttaacc
     5341 agtttatccg tggcggcatc gccttcgcca cgcctccggg tacgccactg gcaccgaaag
     5401 cccaggaagg caaacacttc ctgttgcagg aaagtgaacc gaaagagtgg cgtgaatggg
     5461 gaactgcgct tcccaaataa tgcccacctc gagcagatcc ggctgctaac aaagcccgaa
     5521 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     5581 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taattcttga
     5641 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5701 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6061 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6361 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     6421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6541 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     7021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcc
//
