LOCUS       sequence-333953-        7076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA-[LetB PLL6->1] mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7076)
  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 7076)
  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: pBEL2199
FEATURES             Location/Qualifiers
     source          1..7076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     286..303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    318..457
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(383..402)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     543..565
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(559..750)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1259..1278
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1259..1277
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1309..1331
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5292..5310)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5306..5353
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5398..5416)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5484..5588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5589..5657,5658..6449)
                     /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(5807..5826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6620..7076,1..132)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
       61 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      301 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      361 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
      421 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
      481 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
      541 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
      601 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
      661 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
      721 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
      781 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
      841 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
      901 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
      961 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     1021 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     1081 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     1141 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     1201 gatcatgcgc acccgtggcc aggacccaac gctgcccgag atctcgatcc cgcgaaatta
     1261 atacgactca ctatagggag accacaacgg tttccctcta gaaataattt tgtttaactt
     1321 taagaaggag atatacatat ggctcttaac acaccacaaa ttacgccgac aaaaaagata
     1381 acagtgaggg caatcggcga ggaactgccg cgtggtgatt accaacgttg cccgcaatgt
     1441 gacatgctgt ttagcctgcc cgagataaat tctcatcaaa gtgcctattg tccgcgctgt
     1501 caggcaaaaa ttcgtgacgg gcgcgactgg tcgctaacgc gcctggcggc aatggccttc
     1561 accatgctgt tgcttatgcc gtttgcctgg ggcgaaccgc tgttgcatat ctggctgtta
     1621 ggcattcgta ttgacgccaa cgttatgcaa ggcatctggc aaatgaccaa acagggcgat
     1681 gcgataacgg ggtcgatggt ctttttctgc gttattggtg ccccgctcat tctggtgacc
     1741 tccatagctt atttatggtt tggtaaccga ctgggaatga atttacgtcc ggtactgctg
     1801 atgcttgagc gacttaaaga gtgggtaatg ctcgatatct acctggtcgg cattggcgtt
     1861 gcttctataa aggtacagga ttatgcccat atccaggctg gtgtcggctt gttctctttt
     1921 gtggcgttgg tgattttaac gacggtgacg ttgtcacatc ttaatgtcga agaactgtgg
     1981 gagcgatttt atccgcagcg ccccgctacg cgtagggacg agaaacttcg tgtctgtctt
     2041 gggtgccatt ttaccggcta tccagatcag cgtggtcgct gcccgcgttg ccatatcccg
     2101 ctacgcctgc gtcgccgtca tagtctgcaa aaatgctggg cggcgctgtt agcgtcaatc
     2161 gttttattgt tacctgccaa cctgttgcct atttctatca tttatctgaa cggaggacgg
     2221 caggaagata caattctttc cggaattatg tcgctggcaa gtagcaacat tgcggttgca
     2281 ggaatcgtgt ttatcgccag tattctggta ccgtttacta aagtgatcgt catgttcact
     2341 ttactgttga gcattcattt taaatgccag caaggtttac gcacacgcat tctgttactg
     2401 cggatggtga cctggattgg tcgctggtcg atgctcgacc tgtttgtcat atctttaacc
     2461 atgtcgctga ttaatcgcga tcagatcctc gcttttacta tgggaccggc tgcgttttat
     2521 ttcggcgcag cggtaatttt gactattctt gctgtggaat ggctggacag ccgcttactt
     2581 tgggatgcac atgagtcagg aaacgcccgc ttcgacgact gaagcgcaga ttaaaaataa
     2641 acgccgtatc tcacctttct ggctgctgcc gttcatcgcg ctaatgattg ccagttggct
     2701 gatttgggac agttatcagg accggggtaa taccgtcacc atcgacttta tgtcggcgga
     2761 tggtattgtt ccaggccgta cgcctgttcg ttatcagggc gttgaagtcg gaacagtgca
     2821 ggatatcagc ctcagcgacg atcttcgtaa gattgaagtc aaggtcagca tcaagtccga
     2881 tatgaaagat gcgctgcgcg aagagactca gttctggctg gtgacgccaa aagcatcgtt
     2941 ggcaggtgtc tccgggctgg acgccctcgt cggtgggaac tatatcggca tgatgccggg
     3001 taaaggtaaa gagcaggatc actttgtcgc actcgatacc caaccgaaat atcggctgga
     3061 caatggcgat ctgatgatcc acctgcaagc ccccgatctc ggttcgctga acagcggttc
     3121 attggtctat ttccgcaaga tcccggtggg aaaagtctac gactatgcca tcaatcccaa
     3181 caagcaaggc gtggtgattg atgtcctgat cgagcggcgt tttaccgatc tggtgaaaaa
     3241 aggtagccgt ttctggaacg tttccggcgt tgatgccaac gtcagtatca gtggcgcgaa
     3301 ggtgaaactg gaaagcctgg cggcactggt taacggtgcg attgccttcg attcaccaga
     3361 agagtcgaaa cctgccgagg cggaagatac ctttggtctg tatgaagatc tggcccacag
     3421 ccagcgtggc gtaataataa aactggaact gccgagtggg gccggattaa ccgccgactc
     3481 gacgccgtta atgtatcagg ggctggaagt cggacagctg actaaactgg atttaaatcc
     3541 tggtggtaaa gtcacagggg aaatgaccgt tgatcccagc gtcgttaccc tgcttcgtga
     3601 aaatacccgc atcgaattac gcaacccgaa attatccctt agcgatgcta atctcagcgc
     3661 cctgctgacc ggcaaaacct tcgagctggt gcccggcgat ggcgagccac gcaaagagtt
     3721 cgttgttgtg ccaggcgaaa aagcactgct gcatgaacct gatgttctga cgctgaccct
     3781 gaccgcgccg gaaagttacg gtattgatgc gggtcagccg ctcattcttc acggcgtgca
     3841 ggtaggccag gttatcgatc gtaaactcac cagcaaaggc gtcaccttta ccgtcgccat
     3901 cgagcctcag catcgcgaac tggtaaaagg cgatagcaaa tttgtcgtca acagccgtgt
     3961 cgatgtgaag gtggggctgg atggcgttga gtttctcggt gccagcgcct cagaatggat
     4021 caatggcggg atacgtattc tgccgggcga taaaggtgag atgaaagcca gctatccact
     4081 gtatgccaat ttggaaaaag cgctggagaa cagccttagc gatttaccca ccacaaccgt
     4141 gagtttgagt gcagagacgc tgccggatgt gcaggcagga tcggtagtgc tgtaccgtaa
     4201 atttgaagtt ggtgaagtga ttaccgtgcg tccgcgagct aacgcgtttg atatcgatct
     4261 gcatattaag ccggagtatc gcaaccttct gaccagcaat agcgtgttct gggcagaagg
     4321 cggggcgaaa gttcagctga atggtagtgg tctgaccgta caggcatccc cgctctccag
     4381 agcattaaag ggagccatta gcttcgacaa cctcagcggt gccagcgcca gtcagcgtaa
     4441 aggcgataag cgtattctgt atgcttccga aacagcggcc cgtgcggttg gcgggcagat
     4501 tacgcttcac gctttcgatg ccggaaaact ggcggtcggg atgccaattc gctatctcgg
     4561 tattgatatc gggcaaatcc agacgctgga tctgattacc gcacgcaatg aagtacaggc
     4621 aaaggcggta ctttacccgg aatatgtcca gacctttgct cgcggtggta cgcgcttctc
     4681 agtggtcacg ccaaaagcat cgttggcagg tgtctccggg ctggacgccc tcgtcggtgg
     4741 gaactatatc aacgtcgaac caggccgggg caatcctcgc cgcgactttg aattacaaga
     4801 ggccaccatt actgattcgc gttacctgga tggcttaagc attattgttg aagcgccgga
     4861 agccggttcg ttaggcatcg gtacgcctgt gctgttccgt ggtctggaag tcggtacggt
     4921 tacaggaatg acgctgggga cattgtcaga tcgcgtgatg attgcgatgc gcatcagtaa
     4981 acgctatcaa cacctggtgc gtaacaattc cgtcttctgg ttggcatcgg gttacagtct
     5041 ggactttggt ctgacgggcg gcgtagtgaa aaccggcacc tttaaccagt ttatccgtgg
     5101 cggcatcgcc ttcgccacgc ctccgggtac gccactggca ccgaaagccc aggaaggcaa
     5161 acacttcctg ttgcaggaaa gtgaaccgaa agagtggcgt gaatggggaa ctgcgcttcc
     5221 caaataatgc ccacctcgag cagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     5281 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     5341 gaggggtttt ttgctgaaag gaggaactat atccggataa ttcttgaaga cgaaagggcc
     5401 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5461 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5521 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5581 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5701 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5761 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5821 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5881 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6001 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6121 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6181 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6301 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6421 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6481 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6541 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6601 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6661 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6721 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6781 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6841 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6901 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6961 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7021 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtc
//
