LOCUS       sequence-333952-        6700 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 4 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  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 6700)
  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: pBEL2119
FEATURES             Location/Qualifiers
     source          1..6700
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(131..153)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    239..378
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     294..313
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(393..410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(564..1152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(644..663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1323..2114,2115..2183))
                     /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     1946..1965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2184..2288)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2356..2374
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2419..2466)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2462..2480
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             6065..6087
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6118..6137)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6119..6137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(6646..6700,1..137)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg
       61 aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg
      121 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
      181 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
      241 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
      301 agagcagatt gtactgagag tgcaccattg cggtgtgaaa taccgcacag atgcgtaagg
      361 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      421 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      961 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1321 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1381 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1441 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1501 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1561 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1621 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1681 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1741 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1801 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1861 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1921 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     1981 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2041 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2101 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2161 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2221 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2281 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     2341 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attatccgga
     2401 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
     2461 tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
     2521 agccggatct gctcaggtgg gcattatttg ggaagcgcag ttccccattc acgccactct
     2581 ttcggttcac tttcctgcaa caggaagtgt ttgccttcct gggctttcgg tgccagtggc
     2641 gtacccggag gcgtggcgaa ggcgatgccg ccacggataa actggttaaa ggtgccggtt
     2701 ttcactacgc cgcccgtcag accaaagtcc agactgtaac ccgatgccaa ccagaacacg
     2761 gaattgttac gcaccaggtg ttgatagcgt ttactgatgc gcatcgcaat catcacgcga
     2821 tctgacaatg tccccagcgt cattcctgta accgtaccga cttccagacc acggaacagc
     2881 acaggcgtac cgatgcctaa cgaaccggct tccggcgctt caacaataat gcttaagcca
     2941 tccaggtaac gcgaatcagt aatggtggcc tcttgtaatt caaagtcgcg gcgaggattg
     3001 ccccggcctg gttcgacgtt gatatacggc tggaggatag tatcaagatg ctcaacgcca
     3061 gctgccgaaa tttgcggtgt gaccactgag aagcgcgtac caccgcgagc aaaggtctgg
     3121 acatattccg ggtaaagtac cgcctttgcc tgtacttcat tgcgtgcggt aatcagatcc
     3181 agcgtctgga tttgcccgat atcaataccg agatagcgaa ttggcatccc gaccgccagt
     3241 tttccggcat cgaaagcgtg aagcgtaatc tgcccgccaa ccgcacgggc cgctgtttcg
     3301 gaagcataca gaatacgctt atcgccttta cgctgactgg cgctggcacc gctgaggttg
     3361 tcgaagctaa tggctccctt taatgctctg gagagcgggg atgcctgtac ggtcagacca
     3421 ctaccattca gctgaacttt cgccccgcct tctgcccaga acacgctatt gctggtcaga
     3481 aggttgcgat actccggctt aatatgcaga tcgatatcaa acgcgttagc tcgcggacgc
     3541 acggtaatca cttcaccaac ttcaaattta cggtacagca ctaccgatcc tgcctgcaca
     3601 tccggcagcg tctctgcact caaactcacg gtatcaggtt catgcagcag tgctttttcg
     3661 cctggcacaa caacgaactc tttgcgtggc tcgccatcgc cgggcaccag ctcgaaggtt
     3721 ttgccggtca gcagggcgct gagattagca tcgctaaggg ataatttcgg gttgcgtaat
     3781 tcgatgcggg tattttcacg aagcagggta acgacgctgg gatcaacggt catttcccct
     3841 gtgactttac caccaggatt taaatccagt ttagtcagct gtccgacttc cagcccctga
     3901 tacattaacg gcgtcgagtc ggcggttaat ccggccccac tcggcagttc cagttttatt
     3961 attacgccac gctggctgtg ggccagatct tcatacagac caaaggtatc ttccgcctcg
     4021 gcaggtttcg actcttctgg tgaatcgaag gcaatcgcac cgttaaccag tgccgccagg
     4081 ctttccagtt tcaccttcgc gccactgata ctgacgttgg catcaacgcc ggaaacgttc
     4141 cagaaacggc tacctttttt caccagatcg gtaaaacgcc gctcgatcag gacatcaatc
     4201 accacgcctt gcttgttggg attgatggca tagtcgtaga cttttcccac cgggatcttg
     4261 cggaaataga ccaatgaacc gctgttcagc gaaccgagat cgggggcttg caggtggatc
     4321 atcagatcgc cattgtccag ccgatatttc ggttgggtat cgagtgcgac aaagtgatcc
     4381 tgctctttac ctttacccgg catcatgccg atatagttcc caccgacgag ggcgtccagc
     4441 ccggagacac ctgccaacga tgcttttggc gtcaccagcc agaactgagt ctcttcgcgc
     4501 agcgcatctt tcatatcgga cttgatgctg accttgactt caatcttacg aagatcgtcg
     4561 ctgaggctga tatcctgcac tgttccgact tcaacgccct gataacgaac aggcgtacgg
     4621 cctggaacaa taccatccgc cgacataaag tcgatggtga cggtattacc ccggtcctga
     4681 taactgtccc aaatcagcca actggcaatc attagcgcga tgaacggcag cagccagaaa
     4741 ggtgagatac ggcgtttatt tttaatctgc gcttcagtcg tcgaagcggg cgtttcctga
     4801 ctcatgtgca tcccaaagta agcggctgtc cagccattcc acagcaagaa tagtcaaaat
     4861 taccgctgcg ccgaaataaa acgcagccgg tcccatagta aaagcgagga tctgatcgcg
     4921 attaatcagc gacatggtta aagatatgac aaacaggtcg agcatcgacc agcgaccaat
     4981 ccaggtcacc atccgcagta acagaatgcg tgtgcgtaaa ccttgctggc atttaaaatg
     5041 aatgctcaac agtaaagtga acatgacgat cactttagta aacgggacca gaatactggc
     5101 gataaacacg attcctgcaa ccgcaatgtt gctacttgcc agcgacataa ttccggaaag
     5161 aattgtatct tcctgccgtc ctccgttcag ataaatgata gaaataggca acaggttggc
     5221 aggtaacaat aaaacgattg acgctaacag cgccgcccag catttttgca gactatgacg
     5281 gcgacgcagg cgtagcggga tatggcaacg cgggcagcga ccacgctgat ctggatagcc
     5341 ggtaaaatgg caccctaggc agacacgaag tttctcgtcc ctacgcgtag cggggcgctg
     5401 cggataaaat cgctcccaca gttcttcgac attaagatgt gacaacgtca ccgtcgttaa
     5461 aatcaccaac gccacaaaag agaacaagcc gacaccagcc tggatatggg cataatcctg
     5521 tacctttata gaagcaacgc caatgccgac caggtagata tcgagcatta cccactcttt
     5581 aagtcgctca agcatcagca gtaccggacg taaattcatt cccagtcggt taccaaacca
     5641 taaataagct atggaggtca ccagaatgag cggggcacca ataacgcaga aaaagaccat
     5701 cgaccccgtt atcgcatcgc cctgtttggt catttgccag atgccttgca taacgttggc
     5761 gtcaatacga atgcctaaca gccagatatg caacagcggt tcgccccagg caaacggcat
     5821 aagcaacagc atggtgaagg ccattgccgc caggcgcgtt agcgaccagt cgcgcccgtc
     5881 acgaattttt gcctgacagc gcggacaata ggcactttga tgagaattta tctcgggcag
     5941 gctaaacagc atgtcacatt gcgggcaacg ttggtaatca ccacgcggca gttcctcgcc
     6001 gattgccctc actgttatct tttttgtcgg cgtaatttgt ggtgtgttaa gagccatatg
     6061 tatatctcct tcttaaagtt aaacaaaatt atttctagag ggaaaccgtt gtggtctccc
     6121 tatagtgagt cgtattaatt tcgcgggatc gagatctcgg gcagcgttgg gtcctggcca
     6181 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     6241 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     6301 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     6361 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     6421 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     6481 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     6541 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     6601 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     6661 aaaaccgccc ttaacatggc ccgctttatc agaagccaga
//
