LOCUS       sequence-333945-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 3 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  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 6737)
  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: pBEL2118
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..28
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     262..279
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    294..433
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(359..378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     519..541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(535..726)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1235..1254
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1235..1253
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1285..1307
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4929..4947)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4943..4990
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5035..5053)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5121..5225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5226..5294,5295..6086)
                     /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(5444..5463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6257..6737,1..108)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
       61 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      121 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      181 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
      241 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
      301 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
      361 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
      421 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
      481 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
      541 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
      601 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
      661 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
      721 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
      781 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
      841 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
      901 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
      961 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     1021 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     1081 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     1141 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     1201 cccaacgctg cccgagatct cgatcccgcg aaattaatac gactcactat agggagacca
     1261 caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat acatatggct
     1321 cttaacacac cacaaattac gccgacaaaa aagataacag tgagggcaat cggcgaggaa
     1381 ctgccgcgtg gtgattacca acgttgcccg caatgtgaca tgctgtttag cctgcccgag
     1441 ataaattctc atcaaagtgc ctattgtccg cgctgtcagg caaaaattcg tgacgggcgc
     1501 gactggtcgc taacgcgcct ggcggcaatg gccttcacca tgctgttgct tatgccgttt
     1561 gcctggggcg aaccgctgtt gcatatctgg ctgttaggca ttcgtattga cgccaacgtt
     1621 atgcaaggca tctggcaaat gaccaaacag ggcgatgcga taacggggtc gatggtcttt
     1681 ttctgcgtta ttggtgcccc gctcattctg gtgacctcca tagcttattt atggtttggt
     1741 aaccgactgg gaatgaattt acgtccggta ctgctgatgc ttgagcgact taaagagtgg
     1801 gtaatgctcg atatctacct ggtcggcatt ggcgttgctt ctataaaggt acaggattat
     1861 gcccatatcc aggctggtgt cggcttgttc tcttttgtgg cgttggtgat tttaacgacg
     1921 gtgacgttgt cacatcttaa tgtcgaagaa ctgtgggagc gattttatcc gcagcgcccc
     1981 gctacgcgta gggacgagaa acttcgtgtc tgcctagggt gccattttac cggctatcca
     2041 gatcagcgtg gtcgctgccc gcgttgccat atcccgctac gcctgcgtcg ccgtcatagt
     2101 ctgcaaaaat gctgggcggc gctgttagcg tcaatcgttt tattgttacc tgccaacctg
     2161 ttgcctattt ctatcattta tctgaacgga ggacggcagg aagatacaat tctttccgga
     2221 attatgtcgc tggcaagtag caacattgcg gttgcaggaa tcgtgtttat cgccagtatt
     2281 ctggtcccgt ttactaaagt gatcgtcatg ttcactttac tgttgagcat tcattttaaa
     2341 tgccagcaag gtttacgcac acgcattctg ttactgcgga tggtgacctg gattggtcgc
     2401 tggtcgatgc tcgacctgtt tgtcatatct ttaaccatgt cgctgattaa tcgcgatcag
     2461 atcctcgctt ttactatggg accggctgcg ttttatttcg gcgcagcggt aattttgact
     2521 attcttgctg tggaatggct ggacagccgc ttactttggg atgcacatga gtcaggaaac
     2581 gcccgcttcg acgactgaag cgcagattaa aaataaacgc cgtatctcac ctttctggct
     2641 gctgccgttc atcgcgctaa tgattgccag ttggctgatt tgggacagtt atcaggaccg
     2701 gggtaatacc gtcaccatcg actttatgtc ggcggatggt attgttccag gccgtacgcc
     2761 tgttcgttat cagggcgttg aagtcggaac agtgcaggat atcagcctca gcgacgatct
     2821 tcgtaagatt gaagtcaagg tcagcatcaa gtccgatatg aaagatgcgc tgcgcgaaga
     2881 gactcagttc tggctggtga cgccaaaagc atcgttggca ggtgtctccg ggctggacgc
     2941 cctcgtcggt gggaactata tcggcatgat gccgggtaaa ggtaaagagc aggatcactt
     3001 tgtcgcactc gatacccaac cgaaatatcg gctggacaat ggcgatctga tgatccacct
     3061 gcaagccccc gatctcggct cgctgaacag cggttcattg gtctatttcc gcaagatccc
     3121 ggtgggaaaa gtctacgact atgccatcaa tcccaacaag caaggcgtgg tgattgatgt
     3181 cctgatcgag cggcgtttta ccgatctggt gaaaaaaggt agccgtttct ggaacgtttc
     3241 cggcgttgat gccaacgtca gtatcagtgg cgcgaaggtg aaactggaaa gcctggcggc
     3301 actggttaac ggtgcgattg ccttcgattc accagaagag tcgaaacctg ccgaggcgga
     3361 agataccttt ggtctgtatg aagatctggc ccacagccag cgtgttctga cgctgaccct
     3421 gaccgcgccg gaaagttacg gtattgatgc gggtcagccg ctcattcttc acggcgtgca
     3481 ggtaggccag gttatcgatc gtaaactcac cagcaaaggc gtcaccttta ccgtcgccat
     3541 cgagcctcag catcgcgaac tggtaaaagg cgatagcaaa tttgtcgtca acagccgtgt
     3601 cgatgtgaag gtggggctgg atggcgttga gtttctcggt gccagcgcct cagaatggat
     3661 caatggcggg atacgtattc tgccgggcga taaaggtgag atgaaagcca gctatccact
     3721 gtatgccaat ttggaaaaag cgctggagaa cagccttagc gatttaccca ccacaaccgt
     3781 gagtttgagt gcagagacgc tgccggatgt gcaggcagga tcggtagtgc tgtaccgtaa
     3841 atttgaagtt ggtgaagtga ttaccgtgcg tccgcgagct aacgcgtttg atatcgatct
     3901 gcatattaag ccggagtatc gcaaccttct gaccagcaat agcgtgttct gggcagaagg
     3961 cggggcgaaa gttcagctga atggtagtgg tctgaccgta caggcatccc cgctctccag
     4021 agcattaaag ggagccatta gcttcgacaa cctcagcggt gccagcgcca gtcagcgtaa
     4081 aggcgataag cgtattctgt atgcttccga aacagcggcc cgtgcggttg gcgggcagat
     4141 tacgcttcac gctttcgatg ccggaaaact ggcggtcggg atgccaattc gctatctcgg
     4201 tattgatatc gggcaaatcc agacgctgga tctgattacc gcacgcaatg aagtacaggc
     4261 aaaggcggta ctttacccgg aatatgtcca gacctttgct cgcggtggta cgcgcttctc
     4321 agtggtcaca ccgcaaattt cggcagctgg cgttgagcat cttgatacta tcctccagcc
     4381 gtatatcaac gtcgaaccag gccggggcaa tcctcgccgc gactttgaat tacaagaggc
     4441 caccattact gattcgcgtt acctggatgg cttaagcatt attgttgaag cgccggaagc
     4501 cggttcgtta ggcatcggta cgcctgtgct gttccgtggt ctggaagtcg gtacggttac
     4561 aggaatgacg ctggggacat tgtcagatcg cgtgatgatt gcgatgcgca tcagtaaacg
     4621 ctatcaacac ctggtgcgta acaattccgt gttctggttg gcatcgggtt acagtctgga
     4681 ctttggtctg acgggcggcg tagtgaaaac cggcaccttt aaccagttta tccgtggcgg
     4741 catcgccttc gccacgcctc cgggtacgcc actggcaccg aaagcccagg aaggcaaaca
     4801 cttcctgttg caggaaagtg aaccgaaaga gtggcgtgaa tggggaactg cgcttcccaa
     4861 ataatgccca cctcgagcag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     4921 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     4981 gggttttttg ctgaaaggag gaactatatc cggataattc ttgaagacga aagggcctcg
     5041 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5101 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5221 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5341 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5401 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5461 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5521 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5641 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5761 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5821 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5941 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6061 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6721 gagagcgcac gagggag
//
