LOCUS       sequence-334330-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with LetB mutant L123N.
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: pBEL2343
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3981..3999)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3995..4042
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4087..4105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4173..4277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4278..4346,4347..5138)
                     /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(4496..4515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5309..5897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5798..5817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6051..6068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6083..6222
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6148..6167)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6308..6330
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6324..6515)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     7024..7043
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7024..7042
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atggctctta acacaccaca aattacgccg
       61 acaaaaaaga taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt
      121 tgcccgcaat gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat
      181 tgtccgcgct gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg
      241 gcaatggcct tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat
      301 atctggctgt taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc
      361 aaacagggcg atgcgataac ggggtcgatg gtctttttct gcgttattgg tgccccgctc
      421 attctggtga cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt
      481 ccggtactgc tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc
      541 ggcattggcg ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc
      601 ttgttctctt ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc
      661 gaagaactgt gggagcgatt ttatccgcag cgccccgcta cgcgtaggga cgagaaactt
      721 cgtgtctgtc ttgggtgcca ttttaccggc tatccagatc agcgtggtcg ctgcccgcgt
      781 tgccatatcc cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg ggcggcgctg
      841 ttagcgtcaa tcgttttatt gttacctgcc aacctgttgc ctatttctat catttatctg
      901 aacggaggac ggcaggaaga tacaattctt tccggaatta tgtcgctggc aagtagcaac
      961 attgcggttg caggaatcgt gtttatcgcc agtattctgg taccgtttac taaagtgatc
     1021 gtcatgttca ctttactgtt gagcattcat tttaaatgcc agcaaggttt acgcacacgc
     1081 attctgttac tgcggatggt gacctggatt ggtcgctggt cgatgctcga cctgtttgtc
     1141 atatctttaa ccatgtcgct gattaatcgc gatcagatcc tcgcttttac tatgggaccg
     1201 gctgcgtttt atttcggcgc agcggtaatt ttgactattc ttgctgtgga atggctggac
     1261 agccgcttac tttgggatgc acatgagtca ggaaacgccc gcttcgacga ctgaagcgca
     1321 gattaaaaat aaacgccgta tctcaccttt ctggctgctg ccgttcatcg cgctaatgat
     1381 tgccagttgg ctgatttggg acagttatca ggaccggggt aataccgtca ccatcgactt
     1441 tatgtcggcg gatggtattg ttccaggccg tacgcctgtt cgttatcagg gcgttgaagt
     1501 cggaacagtg caggatatca gcctcagcga cgatcttcgt aagattgaag tcaaggtcag
     1561 catcaagtcc gatatgaaag atgcgctgcg cgaagagact cagttctggc tggtgacgcc
     1621 aaaagcatcg ttggcaggtg tctccgggaa tgacgccctc gtcggtggga actatatcgg
     1681 catgatgccg ggtaaaggta aagagcagga tcactttgtc gcactcgata cccaaccgaa
     1741 atatcggctg gacaatggcg atctgatgat ccacctgcaa gcccccgatc tcggttcgct
     1801 gaacagcggt tcattggtct atttccgcaa gatcccggtg ggaaaagtct acgactatgc
     1861 catcaatccc aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc gttttaccga
     1921 tctggtgaaa aaaggtagcc gtttctggaa cgtttccggc gttgatgcca acgtcagtat
     1981 cagtggcgcg aaggtgaaac tggaaagcct ggcggcactg gttaacggtg cgattgcctt
     2041 cgattcacca gaagagtcga aacctgccga ggcggaagat acctttggtc tgtatgaaga
     2101 tctggcccac agccagcgtg gcgtaataat aaaactggaa ctgccgagtg gggccggatt
     2161 aaccgccgac tcgacgccgt taatgtatca ggggctggaa gtcggacagc tgactaaact
     2221 ggatttaaat cctggtggta aagtcacagg ggaaatgacc gttgatccca gcgtcgttac
     2281 cctgcttcgt gaaaataccc gcatcgaatt acgcaacccg aaattatccc ttagcgatgc
     2341 taatctcagc gccctgctga ccggcaaaac cttcgagctg gtgcccggcg atggcgagcc
     2401 acgcaaagag ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac ctgatgttct
     2461 gacgctgacc ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc cgctcattct
     2521 tcacggcgtg caggtaggcc aggttatcga tcgtaaactc accagcaaag gcgtcacctt
     2581 taccgtcgcc atcgagcctc agcatcgcga actggtaaaa ggcgatagca aatttgtcgt
     2641 caacagccgt gtcgatgtga aggtggggct ggatggcgtt gagtttctcg gtgccagcgc
     2701 ctcagaatgg atcaatggcg ggatacgtat tctgccgggc gataaaggtg agatgaaagc
     2761 cagctatcca ctgtatgcca atttggaaaa agcgctggag aacagcctta gcgatttacc
     2821 caccacaacc gtgagtttga gtgcagagac gctgccggat gtgcaggcag gatcggtagt
     2881 gctgtaccgt aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag ctaacgcgtt
     2941 tgatatcgat ctgcatatta agccggagta tcgcaacctt ctgaccagca atagcgtgtt
     3001 ctgggcagaa ggcggggcga aagttcagct gaatggtagt ggtctgaccg tacaggcatc
     3061 cccgctctcc agagcattaa agggagccat tagcttcgac aacctcagcg gtgccagcgc
     3121 cagtcagcgt aaaggcgata agcgtattct gtatgcttcc gaaacagcgg cccgtgcggt
     3181 tggcgggcag attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat
     3241 tcgctatctc ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa
     3301 tgaagtacag gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg
     3361 tacgcgcttc tcagtggtca caccgcaaat ttcggcagct ggcgttgagc atcttgatac
     3421 tatcctccag ccgtatatca acgtcgaacc aggccggggc aatcctcgcc gcgactttga
     3481 attacaagag gccaccatta ctgattcgcg ttacctggat ggcttaagca ttattgttga
     3541 agcgccggaa gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt
     3601 cggtacggtt acaggaatga cgctggggac attgtcagat cgcgtgatga ttgcgatgcg
     3661 catcagtaaa cgctatcaac acctggtgcg taacaattcc gtcttctggt tggcatcggg
     3721 ttacagtctg gactttggtc tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt
     3781 tatccgtggc ggcatcgcct tcgccacgcc tccgggtacg ccactggcac cgaaagccca
     3841 ggaaggcaaa cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac
     3901 tgcgcttccc aaataatgcc cacctcgagc agatccggct gctaacaaag cccgaaagga
     3961 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     4021 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac
     4081 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4141 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4501 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     4561 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4621 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4681 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4741 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4801 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4861 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4921 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4981 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5041 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5101 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5941 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6001 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6061 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6121 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6181 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     6241 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6301 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     6361 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     6421 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     6481 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     6541 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     6601 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     6661 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     6721 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     6781 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     6841 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     6901 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     6961 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga
     7021 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aat
//
