LOCUS       sequence-334327-        7097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA-[LetB PLL6->4] mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7097)
  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 7097)
  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: pBEL2255
FEATURES             Location/Qualifiers
     source          1..7097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      125..713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     614..633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     867..884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    899..1038
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(964..983)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1124..1146
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1140..1331)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1840..1859
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1840..1858
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1890..1912
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5894..5912)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5908..5955
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6000..6018)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6086..6190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6191..6259,6260..7051)
                     /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(6409..6428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
       61 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
      121 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      181 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      241 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
      301 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      361 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      421 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      481 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      541 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      601 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      661 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      721 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      781 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      841 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      901 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
      961 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     1021 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1081 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1141 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     1201 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     1261 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     1321 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     1381 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     1441 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     1501 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     1561 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     1621 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     1681 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     1741 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     1801 caggacccaa cgctgcccga gatctcgatc ccgcgaaatt aatacgactc actataggga
     1861 gaccacaacg gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata
     1921 tggctcttaa cacaccacaa attacgccga caaaaaagat aacagtgagg gcaatcggcg
     1981 aggaactgcc gcgtggtgat taccaacgtt gcccgcaatg tgacatgctg tttagcctgc
     2041 ccgagataaa ttctcatcaa agtgcctatt gtccgcgctg tcaggcaaaa attcgtgacg
     2101 ggcgcgactg gtcgctaacg cgcctggcgg caatggcctt caccatgctg ttgcttatgc
     2161 cgtttgcctg gggcgaaccg ctgttgcata tctggctgtt aggcattcgt attgacgcca
     2221 acgttatgca aggcatctgg caaatgacca aacagggcga tgcgataacg gggtcgatgg
     2281 tctttttctg cgttattggt gccccgctca ttctggtgac ctccatagct tatttatggt
     2341 ttggtaaccg actgggaatg aatttacgtc cggtactgct gatgcttgag cgacttaaag
     2401 agtgggtaat gctcgatatc tacctggtcg gcattggcgt tgcttctata aaggtacagg
     2461 attatgccca tatccaggct ggtgtcggct tgttctcttt tgtggcgttg gtgattttaa
     2521 cgacggtgac gttgtcacat cttaatgtcg aagaactgtg ggagcgattt tatccgcagc
     2581 gccccgctac gcgtagggac gagaaacttc gtgtctgtct tgggtgccat tttaccggct
     2641 atccagatca gcgtggtcgc tgcccgcgtt gccatatccc gctacgcctg cgtcgccgtc
     2701 atagtctgca aaaatgctgg gcggcgctgt tagcgtcaat cgttttattg ttacctgcca
     2761 acctgttgcc tatttctatc atttatctga acggaggacg gcaggaagat acaattcttt
     2821 ccggaattat gtcgctggca agtagcaaca ttgcggttgc aggaatcgtg tttatcgcca
     2881 gtattctggt accgtttact aaagtgatcg tcatgttcac tttactgttg agcattcatt
     2941 ttaaatgcca gcaaggttta cgcacacgca ttctgttact gcggatggtg acctggattg
     3001 gtcgctggtc gatgctcgac ctgtttgtca tatctttaac catgtcgctg attaatcgcg
     3061 atcagatcct cgcttttact atgggaccgg ctgcgtttta tttcggcgca gcggtaattt
     3121 tgactattct tgctgtggaa tggctggaca gccgcttact ttgggatgca catgagtcag
     3181 gaaacgcccg cttcgacgac tgaagcgcag attaaaaata aacgccgtat ctcacctttc
     3241 tggctgctgc cgttcatcgc gctaatgatt gccagttggc tgatttggga cagttatcag
     3301 gaccggggta ataccgtcac catcgacttt atgtcggcgg atggtattgt tccaggccgt
     3361 acgcctgttc gttatcaggg cgttgaagtc ggaacagtgc aggatatcag cctcagcgac
     3421 gatcttcgta agattgaagt caaggtcagc atcaagtccg atatgaaaga tgcgctgcgc
     3481 gaagagactc agttctggct ggtgacgcca aaagcatcgt tggcaggtgt ctccgggctg
     3541 gacgccctcg tcggtgggaa ctatatcggc atgatgccgg gtaaaggtaa agagcaggat
     3601 cactttgtcg cactcgatac ccaaccgaaa tatcggctgg acaatggcga tctgatgatc
     3661 cacctgcaag cccccgatct cggttcgctg aacagcggtt cattggtcta tttccgcaag
     3721 atcccggtgg gaaaagtcta cgactatgcc atcaatccca acaagcaagg cgtggtgatt
     3781 gatgtcctga tcgagcggcg ttttaccgat ctggtgaaaa aaggtagccg tttctggaac
     3841 gtttccggcg ttgatgccaa cgtcagtatc agtggcgcga aggtgaaact ggaaagcctg
     3901 gcggcactgg ttaacggtgc gattgccttc gattcaccag aagagtcgaa acctgccgag
     3961 gcggaagata cctttggtct gtatgaagat ctggcccaca gccagcgtgg cgtaataata
     4021 aaactggaac tgccgagtgg ggccggatta accgccgact cgacgccgtt aatgtatcag
     4081 gggctggaag tcggacagct gactaaactg gatttaaatc ctggtggtaa agtcacaggg
     4141 gaaatgaccg ttgatcccag cgtcgttacc ctgcttcgtg aaaatacccg catcgaatta
     4201 cgcaacccga aattatccct tagcgatgct aatctcagcg ccctgctgac cggcaaaacc
     4261 ttcgagctgg tgcccggcga tggcgagcca cgcaaagagt tcgttgttgt gccaggcgaa
     4321 aaagcactgc tgcatgaacc tgatgttctg acgctgaccc tgaccgcgcc ggaaagttac
     4381 ggtattgatg cgggtcagcc gctcattctt cacggcgtgc aggtaggcca ggttatcgat
     4441 cgtaaactca ccagcaaagg cgtcaccttt accgtcgcca tcgagcctca gcatcgcgaa
     4501 ctggtaaaag gcgatagcaa atttgtcgtc aacagccgtg tcgatgtgaa ggtggggctg
     4561 gatggcgttg agtttctcgg tgccagcgcc tcagaatgga tcaatggcgg gatacgtatt
     4621 ctgccgggcg ataaaggtga gatgaaagcc agctatccac tgtatgccaa tttggaaaaa
     4681 gcgctggaga acagccttag cgatttaccc accacaaccg tgagtttgag tgcagagacg
     4741 ctgccggatg tgcaggcagg atcggtagtg ctgtaccgta aatttgaagt tggtgaagtg
     4801 attaccgtgc gtccgcgagc taacgcgttt gatatcgatc tgcatattaa gccggagtat
     4861 cgcaaccttc tgaccagcaa tagcgtgttc tgggcagaag gcggggcgaa agttcagctg
     4921 aatggtagtg gtctgaccgt acaggcatcc ccgctctcca gagcattaaa gggagccatt
     4981 agcttcgaca acctcagcgg tgccagcgcc agtcagcgta aaggcgataa gcgtattctg
     5041 tatgcttccg aaacagcggc ccgtgcggtt ggcgggcaga ttacgcttca cgctttcgat
     5101 gccggaaaac tggcggtcgg gatgccaatt cgctatctcg gtattgatat cgggcaaatc
     5161 cagacgctgg atctgattac cgcacgcaat gaagtacagg caaaggcggt actttacccg
     5221 gaatatgtcc agacctttgc tcgcggtggt acgcgcttct cagtggtcag ccgtgtcgat
     5281 gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca gcgcctcaga atggatcaat
     5341 ggcccgtata tcaacgtcga accaggccgg ggcaatcctc gccgcgactt tgaattacaa
     5401 gaggccacca ttactgattc gcgttacctg gatggcttaa gcattattgt tgaagcgccg
     5461 gaagccggtt cgttaggcat cggtacgcct gtgctgttcc gtggtctgga agtcggtacg
     5521 gttacaggaa tgacgctggg gacattgtca gatcgcgtga tgattgcgat gcgcatcagt
     5581 aaacgctatc aacacctggt gcgtaacaat tccgtcttct ggttggcatc gggttacagt
     5641 ctggactttg gtctgacggg cggcgtagtg aaaaccggca cctttaacca gtttatccgt
     5701 ggcggcatcg ccttcgccac gcctccgggt acgccactgg caccgaaagc ccaggaaggc
     5761 aaacacttcc tgttgcagga aagtgaaccg aaagagtggc gtgaatgggg aactgcgctt
     5821 cccaaataat gcccacctcg agcagatccg gctgctaaca aagcccgaaa ggaagctgag
     5881 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     5941 ttgaggggtt ttttgctgaa aggaggaact atatccggat aattcttgaa gacgaaaggg
     6001 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6061 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6121 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6181 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6241 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6301 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6361 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6421 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6481 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6541 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6601 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6661 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6721 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6781 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6841 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6901 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6961 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7021 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7081 ttagattgat ttaaaac
//
