LOCUS       sequence-345246-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with 6PLL3-helix.
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: pBEL2409
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(116..135)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      929..1517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1418..1437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1671..1688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1703..1842
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1768..1787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1928..1950
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1944..2135)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2644..2663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2644..2662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2694..2716
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6674..6692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6688..6735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6780..6798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6866..6970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6971..7039,7040..7073,1..758)
                     /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"
ORIGIN
        1 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
       61 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      121 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
      181 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      241 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
      301 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
      361 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
      421 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
      481 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
      541 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
      601 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      661 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      721 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
      781 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
      841 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
      901 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1441 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1501 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1561 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1621 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     1681 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     1741 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     1801 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     1861 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1921 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     1981 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     2041 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     2101 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     2161 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     2221 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     2281 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     2341 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     2401 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     2461 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     2521 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     2581 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatctc gatcccgcga
     2641 aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat aattttgttt
     2701 aactttaaga aggagatata catatggctc ttaacacacc acaaattacg ccgacaaaaa
     2761 agataacagt gagggcaatc ggcgaggaac tgccgcgtgg tgattaccaa cgttgcccgc
     2821 aatgtgacat gctgtttagc ctgcccgaga taaattctca tcaaagtgcc tattgtccgc
     2881 gctgtcaggc aaaaattcgt gacgggcgcg actggtcgct aacgcgcctg gcggcaatgg
     2941 ccttcaccat gctgttgctt atgccgtttg cctggggcga accgctgttg catatctggc
     3001 tgttaggcat tcgtattgac gccaacgtta tgcaaggcat ctggcaaatg accaaacagg
     3061 gcgatgcgat aacggggtcg atggtctttt tctgcgttat tggtgccccg ctcattctgg
     3121 tgacctccat agcttattta tggtttggta accgactggg aatgaattta cgtccggtac
     3181 tgctgatgct tgagcgactt aaagagtggg taatgctcga tatctacctg gtcggcattg
     3241 gcgttgcttc tataaaggta caggattatg cccatatcca ggctggtgtc ggcttgttct
     3301 cttttgtggc gttggtgatt ttaacgacgg tgacgttgtc acatcttaat gtcgaagaac
     3361 tgtgggagcg attttatccg cagcgccccg ctacgcgtag ggacgagaaa cttcgtgtct
     3421 gtcttgggtg ccattttacc ggctatccag atcagcgtgg tcgctgcccg cgttgccata
     3481 tcccgctacg cctgcgtcgc cgtcatagtc tgcaaaaatg ctgggcggcg ctgttagcgt
     3541 caatcgtttt attgttacct gccaacctgt tgcctatttc tatcatttat ctgaacggag
     3601 gacggcagga agatacaatt ctttccggaa ttatgtcgct ggcaagtagc aacattgcgg
     3661 ttgcaggaat cgtgtttatc gccagtattc tggtaccgtt tactaaagtg atcgtcatgt
     3721 tcactttact gttgagcatt cattttaaat gccagcaagg tttacgcaca cgcattctgt
     3781 tactgcggat ggtgacctgg attggtcgct ggtcgatgct cgacctgttt gtcatatctt
     3841 taaccatgtc gctgattaat cgcgatcaga tcctcgcttt tactatggga ccggctgcgt
     3901 tttatttcgg cgcagcggta attttgacta ttcttgctgt ggaatggctg gacagccgct
     3961 tactttggga tgcacatgag tcaggaaacg cccgcttcga cgactgaagc gcagattaaa
     4021 aataaacgcc gtatctcacc tttctggctg ctgccgttca tcgcgctaat gattgccagt
     4081 tggctgattt gggacagtta tcaggaccgg ggtaataccg tcaccatcga ctttatgtcg
     4141 gcggatggta ttgttccagg ccgtacgcct gttcgttatc agggcgttga agtcggaaca
     4201 gtgcaggata tcagcctcag cgacgatctt cgtaagattg aagtcaaggt cagcatcaag
     4261 tccgatatga aagatgcgct gcgcgaagag actcagttct ggctggtgac gccaaaagca
     4321 tcgttggcag gtgtctccgg gctggacgcc ctcgtcggtg ggaactatat cggcatgatg
     4381 ccgggtaaag gtaaagagca ggatcacttt gtcgcactcg atacccaacc gaaatatcgg
     4441 ctggacaatg gcgatctgat gatccacctg caagcccccg atctcggttc gctgaacagc
     4501 ggttcattgg tctatttccg caagatcccg gtgggaaaag tctacgacta tgccatcaat
     4561 cccaacaagc aaggcgtggt gattgatgtc ctgatcgagc ggcgttttac cgatctggtg
     4621 aaaaaaggta gccgtttctg gaacgtttcc ggcgttgatg ccaacgtcag tatcagtggc
     4681 gcgaaggtga aactggaaag cctggcggca ctggttaacg gtgcgattgc cttcgattca
     4741 ccagaagagt cgaaacctgc cgaggcggaa gatacctttg gtctgtatga agatctggcc
     4801 cacagccagc gtggcgtaat aataaaactg gaactgccga gtggggccgg attaaccgcc
     4861 gactcgacgc cgttaatgta tcaggggctg gaagtcggac agctgactaa actggattta
     4921 aatcctggtg gtaaagtcac aggggaaatg accgttgatc ccagcgtcgt taccctgctt
     4981 cgtgaaaata cccgcatcga attacgcaac ccgaaattat cccttagcga tgctaatctc
     5041 agcgccctgc tgaccggcaa aaccttcgag ctggtgcccg gcgatggcga gccacgcaaa
     5101 gagttcgttg ttgtgccagg cgaaaaagca ctgctgcatg aacctgatgt tctgacgctg
     5161 accctgaccg cgccggaaag ttacggtatt gatgcgggtc agccgctcat tcttcacggc
     5221 gtgcaggtag gccaggttat cgatcgtaaa ctcaccagca aaggcgtcac ctttaccgtc
     5281 gccatcgagc ctcagcatcg cgaactggta aaaggcgata gcaaatttgt cgtcaacagc
     5341 cgtgtcgatg tgaaggtggg gctggatggc gttgagtttc tcggtgccag cgcctcagaa
     5401 tggatcaatg gcgggatacg tattctgccg ggcgataaag gtgagatgaa agccagctat
     5461 ccactgtatg ccaatttgga aaaagcgctg gagaacagcc ttagcgattt acccaccaca
     5521 accgtgagtt tgagtgcaga gacgctgccg gatgtgcagg caggatcggt agtgctgtac
     5581 cgtaaatttg aagttggtga agtgattacc gtgcgtccgc gagctaacgc gtttgatatc
     5641 gatctgcata ttaagccgga gtatcgcaac cttctgacca gcaatagcgt gttctgggca
     5701 gaaggcgggg cgaaagttca gctgaatggt agtggtctga ccgtacaggc atccccgctc
     5761 tccagagcat taaagggagc cattagcttc gacaacctca gcggtgccag cgccagtcag
     5821 cgtaaaggcg ataagcgtat tctgtatgct tccgaaacag cggcccgtgc ggttggcggg
     5881 cagattacgc ttcacgcttt cgatgccgga aaactggcgg tcgggatgcc aattcgctat
     5941 ctcggtattg atatcgggca aatccagacg ctggatctga ttaccgcacg caatgaagta
     6001 caggcaaagg cggtacttta cccggaatat gtccagacct ttgctcgcgg tggtacgcgc
     6061 ttctcagtgg tcacaccgca aatttcggca gctggcgttg agcatctcag cgccctgctg
     6121 cagccgtata tcaacgtcga accaggccgg ggcaatcctc gccgcgactt tgaattacaa
     6181 gaggccacca ttactgattc gcgttacctg gatggcttaa gcattattgt tgaagcgccg
     6241 gaagccggtt cgttaggcat cggtacgcct gtgctgttcc gtggtctgga agtcggtacg
     6301 gttacaggaa tgacgctggg gacattgtca gatcgcgtga tgattgcgat gcgcatcagt
     6361 aaacgctatc aacacctggt gcgtaacaat tccgtcttct ggttggcatc gggttacagt
     6421 ctggactttg gtctgacggg cggcgtagtg aaaaccggca cctttaacca gtttatccgt
     6481 ggcggcatcg ccttcgccac gcctccgggt acgccactgg caccgaaagc ccaggaaggc
     6541 aaacacttcc tgttgcagga aagtgaaccg aaagagtggc gtgaatgggg aactgcgctt
     6601 cccaaataat gcccacctcg agcagatccg gctgctaaca aagcccgaaa ggaagctgag
     6661 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     6721 ttgaggggtt ttttgctgaa aggaggaact atatccggat aattcttgaa gacgaaaggg
     6781 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctg
//
