LOCUS       sequence-333957-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA-[LetB PLL6->5] mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  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 7091)
  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: pBEL2201
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..126)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    212..351
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     267..286
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(366..383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(537..1125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(617..636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1296..2087,2088..2156))
                     /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     1919..1938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2157..2261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2329..2347
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2392..2439)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2435..2453
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             6429..6451
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6482..6501)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6483..6501)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(7010..7091,1..110)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc
       61 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      121 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      181 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
      241 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
      301 ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
      361 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      421 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      481 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      541 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      601 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      661 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      721 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      781 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      841 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      901 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      961 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1021 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1081 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1141 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1201 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1261 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1321 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1381 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1441 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1501 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1561 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     1621 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1681 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1741 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1801 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1861 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     1921 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1981 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2041 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2101 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2161 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2221 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2281 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2341 acgaggccct ttcgtcttca agaattatcc ggatatagtt cctcctttca gcaaaaaacc
     2401 cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag cggtggcagc
     2461 agccaactca gcttcctttc gggctttgtt agcagccgga tctgctcgag gtgggcatta
     2521 tttgggaagc gcagttcccc attcacgcca ctctttcggt tcactttcct gcaacaggaa
     2581 gtgtttgcct tcctgggctt tcggtgccag tggcgtaccc ggaggcgtgg cgaaggcgat
     2641 gccgccacgg ataaactggt taaaggtgcc ggttttcact acgccgcccg tcagaccaaa
     2701 gtccagactg taacccgatg ccaaccagaa gacggaattg ttacgcacca ggtgttgata
     2761 gcgtttactg atgcgcatcg caatcatcac gcgatctgac aatgtcccca gcgtcattcc
     2821 tgtaaccgta ccgacttcca gaccacggaa cagcacaggc gtaccgatgc ctaacgaacc
     2881 ggcttccggc gcttcaacaa taatgcttaa gccatccagg taacgcgaat cagtaatggt
     2941 ggcctcttgt aattcaaagt cgcggcgagg attgccccgg cctggttcga cgttgatata
     3001 cggctttaat gctctggaga gcggggatgc ctgtacggtc agaccactac cattcagctg
     3061 aactttcggt gtgaccactg agaagcgcgt accaccgcga gcaaaggtct ggacatattc
     3121 cgggtaaagt accgcctttg cctgtacttc attgcgtgcg gtaatcagat ccagcgtctg
     3181 gatttgcccg atatcaatac cgagatagcg aattggcatc ccgaccgcca gttttccggc
     3241 atcgaaagcg tgaagcgtaa tctgcccgcc aaccgcacgg gccgctgttt cggaagcata
     3301 cagaatacgc ttatcgcctt tacgctgact ggcgctggca ccgctgaggt tgtcgaagct
     3361 aatggctccc tttaatgctc tggagagcgg ggatgcctgt acggtcagac cactaccatt
     3421 cagctgaact ttcgccccgc cttctgccca gaacacgcta ttgctggtca gaaggttgcg
     3481 atactccggc ttaatatgca gatcgatatc aaacgcgtta gctcgcggac gcacggtaat
     3541 cacttcacca acttcaaatt tacggtacag cactaccgat cctgcctgca catccggcag
     3601 cgtctctgca ctcaaactca cggttgtggt gggtaaatcg ctaaggctgt tctccagcgc
     3661 tttttccaaa ttggcataca gtggatagct ggctttcatc tcacctttat cgcccggcag
     3721 aatacgtatc ccgccattga tccattctga ggcgctggca ccgagaaact caacgccatc
     3781 cagccccacc ttcacatcga cacggctgtt gacgacaaat ttgctatcgc cttttaccag
     3841 ttcgcgatgc tgaggctcga tggcgacggt aaaggtgacg cctttgctgg tgagtttacg
     3901 atcgataacc tggcctacct gcacgccgtg aagaatgagc ggctgacccg catcaatacc
     3961 gtaactttcc ggcgcggtca gggtcagcgt cagaacatca ggttcatgca gcagtgcttt
     4021 ttcgcctggc acaacaacga actctttgcg tggctcgcca tcgccgggca ccagctcgaa
     4081 ggttttgccg gtcagcaggg cgctgagatt agcatcgcta agggataatt tcgggttgcg
     4141 taattcgatg cgggtatttt cacgaagcag ggtaacgacg ctgggatcaa cggtcatttc
     4201 ccctgtgact ttaccaccag gatttaaatc cagtttagtc agctgtccga cttccagccc
     4261 ctgatacatt aacggcgtcg agtcggcggt taatccggcc ccactcggca gttccagttt
     4321 tattattacg ccacgctggc tgtgggccag atcttcatac agaccaaagg tatcttccgc
     4381 ctcggcaggt ttcgactctt ctggtgaatc gaaggcaatc gcaccgttaa ccagtgccgc
     4441 caggctttcc agtttcacct tcgcgccact gatactgacg ttggcatcaa cgccggaaac
     4501 gttccagaaa cggctacctt ttttcaccag atcggtaaaa cgccgctcga tcaggacatc
     4561 aatcaccacg ccttgcttgt tgggattgat ggcatagtcg tagacttttc ccaccgggat
     4621 cttgcggaaa tagaccaatg aaccgctgtt cagcgaaccg agatcggggg cttgcaggtg
     4681 gatcatcaga tcgccattgt ccagccgata tttcggttgg gtatcgagtg cgacaaagtg
     4741 atcctgctct ttacctttac ccggcatcat gccgatatag ttcccaccga cgagggcgtc
     4801 cagcccggag acacctgcca acgatgcttt tggcgtcacc agccagaact gagtctcttc
     4861 gcgcagcgca tctttcatat cggacttgat gctgaccttg acttcaatct tacgaagatc
     4921 gtcgctgagg ctgatatcct gcactgttcc gacttcaacg ccctgataac gaacaggcgt
     4981 acggcctgga acaataccat ccgccgacat aaagtcgatg gtgacggtat taccccggtc
     5041 ctgataactg tcccaaatca gccaactggc aatcattagc gcgatgaacg gcagcagcca
     5101 gaaaggtgag atacggcgtt tatttttaat ctgcgcttca gtcgtcgaag cgggcgtttc
     5161 ctgactcatg tgcatcccaa agtaagcggc tgtccagcca ttccacagca agaatagtca
     5221 aaattaccgc tgcgccgaaa taaaacgcag ccggtcccat agtaaaagcg aggatctgat
     5281 cgcgattaat cagcgacatg gttaaagata tgacaaacag gtcgagcatc gaccagcgac
     5341 caatccaggt caccatccgc agtaacagaa tgcgtgtgcg taaaccttgc tggcatttaa
     5401 aatgaatgct caacagtaaa gtgaacatga cgatcacttt agtaaacggt accagaatac
     5461 tggcgataaa cacgattcct gcaaccgcaa tgttgctact tgccagcgac ataattccgg
     5521 aaagaattgt atcttcctgc cgtcctccgt tcagataaat gatagaaata ggcaacaggt
     5581 tggcaggtaa caataaaacg attgacgcta acagcgccgc ccagcatttt tgcagactat
     5641 gacggcgacg caggcgtagc gggatatggc aacgcgggca gcgaccacgc tgatctggat
     5701 agccggtaaa atggcaccca agacagacac gaagtttctc gtccctacgc gtagcggggc
     5761 gctgcggata aaatcgctcc cacagttctt cgacattaag atgtgacaac gtcaccgtcg
     5821 ttaaaatcac caacgccaca aaagagaaca agccgacacc agcctggata tgggcataat
     5881 cctgtacctt tatagaagca acgccaatgc cgaccaggta gatatcgagc attacccact
     5941 ctttaagtcg ctcaagcatc agcagtaccg gacgtaaatt cattcccagt cggttaccaa
     6001 accataaata agctatggag gtcaccagaa tgagcggggc accaataacg cagaaaaaga
     6061 ccatcgaccc cgttatcgca tcgccctgtt tggtcatttg ccagatgcct tgcataacgt
     6121 tggcgtcaat acgaatgcct aacagccaga tatgcaacag cggttcgccc caggcaaacg
     6181 gcataagcaa cagcatggtg aaggccattg ccgccaggcg cgttagcgac cagtcgcgcc
     6241 cgtcacgaat ttttgcctga cagcgcggac aataggcact ttgatgagaa tttatctcgg
     6301 gcaggctaaa cagcatgtca cattgcgggc aacgttggta atcaccacgc ggcagttcct
     6361 cgccgattgc cctcactgtt atcttttttg tcggcgtaat ttgtggtgtg ttaagagcca
     6421 tatgtatatc tccttcttaa agttaaacaa aattatttct agagggaaac cgttgtggtc
     6481 tccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgggcagcg ttgggtcctg
     6541 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     6601 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     6661 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     6721 tctggaaacg cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg
     6781 ctgctggcta ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg
     6841 agtgattttt ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc
     6901 cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat
     6961 cggtatcatt acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca
     7021 ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga
     7081 gaaactcaac g
//
