LOCUS       sequence-345251-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB(I703N).
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: pBEL2415
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     290..309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     543..560
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    575..714
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(640..659)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     800..822
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(816..1007)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1516..1535
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1516..1534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1566..1588
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5546..5564)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5560..5607
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5652..5670)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5738..5842
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5843..5911,5912..6703)
                     /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(6061..6080)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6874..7073,1..389)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
       61 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      421 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      481 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      541 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
      601 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
      661 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
      721 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      781 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      841 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
      901 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
      961 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     1021 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     1081 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     1141 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     1201 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     1261 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     1321 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     1381 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     1441 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatc
     1501 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
     1561 ataattttgt ttaactttaa gaaggagata tacatatggc tcttaacaca ccacaaatta
     1621 cgccgacaaa aaagataaca gtgagggcaa tcggcgagga actgccgcgt ggtgattacc
     1681 aacgttgccc gcaatgtgac atgctgttta gcctgcccga gataaattct catcaaagtg
     1741 cctattgtcc gcgctgtcag gcaaaaattc gtgacgggcg cgactggtcg ctaacgcgcc
     1801 tggcggcaat ggccttcacc atgctgttgc ttatgccgtt tgcctggggc gaaccgctgt
     1861 tgcatatctg gctgttaggc attcgtattg acgccaacgt tatgcaaggc atctggcaaa
     1921 tgaccaaaca gggcgatgcg ataacggggt cgatggtctt tttctgcgtt attggtgccc
     1981 cgctcattct ggtgacctcc atagcttatt tatggtttgg taaccgactg ggaatgaatt
     2041 tacgtccggt actgctgatg cttgagcgac ttaaagagtg ggtaatgctc gatatctacc
     2101 tggtcggcat tggcgttgct tctataaagg tacaggatta tgcccatatc caggctggtg
     2161 tcggcttgtt ctcttttgtg gcgttggtga ttttaacgac ggtgacgttg tcacatctta
     2221 atgtcgaaga actgtgggag cgattttatc cgcagcgccc cgctacgcgt agggacgaga
     2281 aacttcgtgt ctgtcttggg tgccatttta ccggctatcc agatcagcgt ggtcgctgcc
     2341 cgcgttgcca tatcccgcta cgcctgcgtc gccgtcatag tctgcaaaaa tgctgggcgg
     2401 cgctgttagc gtcaatcgtt ttattgttac ctgccaacct gttgcctatt tctatcattt
     2461 atctgaacgg aggacggcag gaagatacaa ttctttccgg aattatgtcg ctggcaagta
     2521 gcaacattgc ggttgcagga atcgtgttta tcgccagtat tctggtaccg tttactaaag
     2581 tgatcgtcat gttcacttta ctgttgagca ttcattttaa atgccagcaa ggtttacgca
     2641 cacgcattct gttactgcgg atggtgacct ggattggtcg ctggtcgatg ctcgacctgt
     2701 ttgtcatatc tttaaccatg tcgctgatta atcgcgatca gatcctcgct tttactatgg
     2761 gaccggctgc gttttatttc ggcgcagcgg taattttgac tattcttgct gtggaatggc
     2821 tggacagccg cttactttgg gatgcacatg agtcaggaaa cgcccgcttc gacgactgaa
     2881 gcgcagatta aaaataaacg ccgtatctca cctttctggc tgctgccgtt catcgcgcta
     2941 atgattgcca gttggctgat ttgggacagt tatcaggacc ggggtaatac cgtcaccatc
     3001 gactttatgt cggcggatgg tattgttcca ggccgtacgc ctgttcgtta tcagggcgtt
     3061 gaagtcggaa cagtgcagga tatcagcctc agcgacgatc ttcgtaagat tgaagtcaag
     3121 gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag agactcagtt ctggctggtg
     3181 acgccaaaag catcgttggc aggtgtctcc gggctggacg ccctcgtcgg tgggaactat
     3241 atcggcatga tgccgggtaa aggtaaagag caggatcact ttgtcgcact cgatacccaa
     3301 ccgaaatatc ggctggacaa tggcgatctg atgatccacc tgcaagcccc cgatctcggt
     3361 tcgctgaaca gcggttcatt ggtctatttc cgcaagatcc cggtgggaaa agtctacgac
     3421 tatgccatca atcccaacaa gcaaggcgtg gtgattgatg tcctgatcga gcggcgtttt
     3481 accgatctgg tgaaaaaagg tagccgtttc tggaacgttt ccggcgttga tgccaacgtc
     3541 agtatcagtg gcgcgaaggt gaaactggaa agcctggcgg cactggttaa cggtgcgatt
     3601 gccttcgatt caccagaaga gtcgaaacct gccgaggcgg aagatacctt tggtctgtat
     3661 gaagatctgg cccacagcca gcgtggcgta ataataaaac tggaactgcc gagtggggcc
     3721 ggattaaccg ccgactcgac gccgttaatg tatcaggggc tggaagtcgg acagctgact
     3781 aaactggatt taaatcctgg tggtaaagtc acaggggaaa tgaccgttga tcccagcgtc
     3841 gttaccctgc ttcgtgaaaa tacccgcatc gaattacgca acccgaaatt atcccttagc
     3901 gatgctaatc tcagcgccct gctgaccggc aaaaccttcg agctggtgcc cggcgatggc
     3961 gagccacgca aagagttcgt tgttgtgcca ggcgaaaaag cactgctgca tgaacctgat
     4021 gttctgacgc tgaccctgac cgcgccggaa agttacggta ttgatgcggg tcagccgctc
     4081 attcttcacg gcgtgcaggt aggccaggtt atcgatcgta aactcaccag caaaggcgtc
     4141 acctttaccg tcgccatcga gcctcagcat cgcgaactgg taaaaggcga tagcaaattt
     4201 gtcgtcaaca gccgtgtcga tgtgaaggtg gggctggatg gcgttgagtt tctcggtgcc
     4261 agcgcctcag aatggatcaa tggcgggata cgtattctgc cgggcgataa aggtgagatg
     4321 aaagccagct atccactgta tgccaatttg gaaaaagcgc tggagaacag ccttagcgat
     4381 ttacccacca caaccgtgag tttgagtgca gagacgctgc cggatgtgca ggcaggatcg
     4441 gtagtgctgt accgtaaatt tgaagttggt gaagtgatta ccgtgcgtcc gcgagctaac
     4501 gcgtttgata tcgatctgca tattaagccg gagtatcgca accttctgac cagcaatagc
     4561 gtgttctggg cagaaggcgg ggcgaaagtt cagctgaatg gtagtggtct gaccgtacag
     4621 gcatccccgc tctccagagc attaaaggga gccattagct tcgacaacct cagcggtgcc
     4681 agcgccagtc agcgtaaagg cgataagcgt attctgtatg cttccgaaac agcggcccgt
     4741 gcggttggcg ggcagattac gcttcacgct ttcgatgccg gaaaactggc ggtcgggatg
     4801 ccaattcgct atctcggtat tgatatcggg caaatccaga cgctggatct gattaccgca
     4861 cgcaatgaag tacaggcaaa ggcggtactt tacccggaat atgtccagac ctttgctcgc
     4921 ggtggtacgc gcttctcagt ggtcacaccg caaaattcgg cagctggcgt tgagcatctt
     4981 gatactatcc tccagccgta tatcaacgtc gaaccaggcc ggggcaatcc tcgccgcgac
     5041 tttgaattac aagaggccac cattactgat tcgcgttacc tggatggctt aagcattatt
     5101 gttgaagcgc cggaagccgg ttcgttaggc atcggtacgc ctgtgctgtt ccgtggtctg
     5161 gaagtcggta cggttacagg aatgacgctg gggacattgt cagatcgcgt gatgattgcg
     5221 atgcgcatca gtaaacgcta tcaacacctg gtgcgtaaca attccgtctt ctggttggca
     5281 tcgggttaca gtctggactt tggtctgacg ggcggcgtag tgaaaaccgg cacctttaac
     5341 cagtttatcc gtggcggcat cgccttcgcc acgcctccgg gtacgccact ggcaccgaaa
     5401 gcccaggaag gcaaacactt cctgttgcag gaaagtgaac cgaaagagtg gcgtgaatgg
     5461 ggaactgcgc ttcccaaata atgcccacct cgagcagatc cggctgctaa caaagcccga
     5521 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     5581 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ataattcttg
     5641 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5701 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5821 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5881 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5941 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6001 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6061 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     6121 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6181 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6241 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6301 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6361 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     6421 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6481 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6541 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6601 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6661 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6721 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6781 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7021 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgc
//
