LOCUS       sequence-333958-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA-[LetB PLL6->7] mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  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 7094)
  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: pBEL2256
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(394..1185,1186..1254))
                     /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     1017..1036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1255..1359)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1427..1445
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1490..1537)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1533..1551
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             5530..5552
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5583..5602)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5584..5602)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6111..6302
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6296..6318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6404..6543
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6459..6478
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6558..6575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6729..7094,1..223))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6809..6828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
       61 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      241 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      301 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      361 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      421 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      481 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      541 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      601 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      661 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt
      721 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      781 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      841 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
      901 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
      961 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa
     1021 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1081 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1141 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1201 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1321 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1381 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     1441 gaggcccttt cgtcttcaag aattatccgg atatagttcc tcctttcagc aaaaaacccc
     1501 tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag
     1561 ccaactcagc ttcctttcgg gctttgttag cagccggatc tgctcgaggt gggcattatt
     1621 tgggaagcgc agttccccat tcacgccact ctttcggttc actttcctgc aacaggaagt
     1681 gtttgccttc ctgggctttc ggtgccagtg gcgtacccgg aggcgtggcg aaggcgatgc
     1741 cgccacggat aaactggtta aaggtgccgg ttttcactac gccgcccgtc agaccaaagt
     1801 ccagactgta acccgatgcc aaccagaaga cggaattgtt acgcaccagg tgttgatagc
     1861 gtttactgat gcgcatcgca atcatcacgc gatctgacaa tgtccccagc gtcattcctg
     1921 taaccgtacc gacttccaga ccacggaaca gcacaggcgt accgatgcct aacgaaccgg
     1981 cttccggcgc ttcaacaata atgcttaagc catccaggta acgcgaatca gtaatggtgg
     2041 cctcttgtaa ttcaaagtcg cggcgaggat tgccccggcc tggttcgacg ttgatatagc
     2101 cacggataaa ctggttaaag gtgccggttt tcactacgcc gcccgtcaga ccaaagtcca
     2161 gactgtaacc cgagaccact gagaagcgcg taccaccgcg agcaaaggtc tggacatatt
     2221 ccgggtaaag taccgccttt gcctgtactt cattgcgtgc ggtaatcaga tccagcgtct
     2281 ggatttgccc gatatcaata ccgagatagc gaattggcat cccgaccgcc agttttccgg
     2341 catcgaaagc gtgaagcgta atctgcccgc caaccgcacg ggccgctgtt tcggaagcat
     2401 acagaatacg cttatcgcct ttacgctgac tggcgctggc accgctgagg ttgtcgaagc
     2461 taatggctcc ctttaatgct ctggagagcg gggatgcctg tacggtcaga ccactaccat
     2521 tcagctgaac tttcgccccg ccttctgccc agaacacgct attgctggtc agaaggttgc
     2581 gatactccgg cttaatatgc agatcgatat caaacgcgtt agctcgcgga cgcacggtaa
     2641 tcacttcacc aacttcaaat ttacggtaca gcactaccga tcctgcctgc acatccggca
     2701 gcgtctctgc actcaaactc acggttgtgg tgggtaaatc gctaaggctg ttctccagcg
     2761 ctttttccaa attggcatac agtggatagc tggctttcat ctcaccttta tcgcccggca
     2821 gaatacgtat cccgccattg atccattctg aggcgctggc accgagaaac tcaacgccat
     2881 ccagccccac cttcacatcg acacggctgt tgacgacaaa tttgctatcg ccttttacca
     2941 gttcgcgatg ctgaggctcg atggcgacgg taaaggtgac gcctttgctg gtgagtttac
     3001 gatcgataac ctggcctacc tgcacgccgt gaagaatgag cggctgaccc gcatcaatac
     3061 cgtaactttc cggcgcggtc agggtcagcg tcagaacatc aggttcatgc agcagtgctt
     3121 tttcgcctgg cacaacaacg aactctttgc gtggctcgcc atcgccgggc accagctcga
     3181 aggttttgcc ggtcagcagg gcgctgagat tagcatcgct aagggataat ttcgggttgc
     3241 gtaattcgat gcgggtattt tcacgaagca gggtaacgac gctgggatca acggtcattt
     3301 cccctgtgac tttaccacca ggatttaaat ccagtttagt cagctgtccg acttccagcc
     3361 cctgatacat taacggcgtc gagtcggcgg ttaatccggc cccactcggc agttccagtt
     3421 ttattattac gccacgctgg ctgtgggcca gatcttcata cagaccaaag gtatcttccg
     3481 cctcggcagg tttcgactct tctggtgaat cgaaggcaat cgcaccgtta accagtgccg
     3541 ccaggctttc cagtttcacc ttcgcgccac tgatactgac gttggcatca acgccggaaa
     3601 cgttccagaa acggctacct tttttcacca gatcggtaaa acgccgctcg atcaggacat
     3661 caatcaccac gccttgcttg ttgggattga tggcatagtc gtagactttt cccaccggga
     3721 tcttgcggaa atagaccaat gaaccgctgt tcagcgaacc gagatcgggg gcttgcaggt
     3781 ggatcatcag atcgccattg tccagccgat atttcggttg ggtatcgagt gcgacaaagt
     3841 gatcctgctc tttaccttta cccggcatca tgccgatata gttcccaccg acgagggcgt
     3901 ccagcccgga gacacctgcc aacgatgctt ttggcgtcac cagccagaac tgagtctctt
     3961 cgcgcagcgc atctttcata tcggacttga tgctgacctt gacttcaatc ttacgaagat
     4021 cgtcgctgag gctgatatcc tgcactgttc cgacttcaac gccctgataa cgaacaggcg
     4081 tacggcctgg aacaatacca tccgccgaca taaagtcgat ggtgacggta ttaccccggt
     4141 cctgataact gtcccaaatc agccaactgg caatcattag cgcgatgaac ggcagcagcc
     4201 agaaaggtga gatacggcgt ttatttttaa tctgcgcttc agtcgtcgaa gcgggcgttt
     4261 cctgactcat gtgcatccca aagtaagcgg ctgtccagcc attccacagc aagaatagtc
     4321 aaaattaccg ctgcgccgaa ataaaacgca gccggtccca tagtaaaagc gaggatctga
     4381 tcgcgattaa tcagcgacat ggttaaagat atgacaaaca ggtcgagcat cgaccagcga
     4441 ccaatccagg tcaccatccg cagtaacaga atgcgtgtgc gtaaaccttg ctggcattta
     4501 aaatgaatgc tcaacagtaa agtgaacatg acgatcactt tagtaaacgg taccagaata
     4561 ctggcgataa acacgattcc tgcaaccgca atgttgctac ttgccagcga cataattccg
     4621 gaaagaattg tatcttcctg ccgtcctccg ttcagataaa tgatagaaat aggcaacagg
     4681 ttggcaggta acaataaaac gattgacgct aacagcgccg cccagcattt ttgcagacta
     4741 tgacggcgac gcaggcgtag cgggatatgg caacgcgggc agcgaccacg ctgatctgga
     4801 tagccggtaa aatggcaccc aagacagaca cgaagtttct cgtccctacg cgtagcgggg
     4861 cgctgcggat aaaatcgctc ccacagttct tcgacattaa gatgtgacaa cgtcaccgtc
     4921 gttaaaatca ccaacgccac aaaagagaac aagccgacac cagcctggat atgggcataa
     4981 tcctgtacct ttatagaagc aacgccaatg ccgaccaggt agatatcgag cattacccac
     5041 tctttaagtc gctcaagcat cagcagtacc ggacgtaaat tcattcccag tcggttacca
     5101 aaccataaat aagctatgga ggtcaccaga atgagcgggg caccaataac gcagaaaaag
     5161 accatcgacc ccgttatcgc atcgccctgt ttggtcattt gccagatgcc ttgcataacg
     5221 ttggcgtcaa tacgaatgcc taacagccag atatgcaaca gcggttcgcc ccaggcaaac
     5281 ggcataagca acagcatggt gaaggccatt gccgccaggc gcgttagcga ccagtcgcgc
     5341 ccgtcacgaa tttttgcctg acagcgcgga caataggcac tttgatgaga atttatctcg
     5401 ggcaggctaa acagcatgtc acattgcggg caacgttggt aatcaccacg cggcagttcc
     5461 tcgccgattg ccctcactgt tatctttttt gtcggcgtaa tttgtggtgt gttaagagcc
     5521 atatgtatat ctccttctta aagttaaaca aaattatttc tagagggaaa ccgttgtggt
     5581 ctccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgggcagc gttgggtcct
     5641 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     5701 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     5761 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     5821 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat
     5881 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct
     5941 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
     6001 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
     6061 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
     6121 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     6181 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     6241 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     6301 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6361 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     6421 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     6481 gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6541 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6601 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6661 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6721 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6781 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6841 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6901 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6961 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7021 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7081 gccactggta acag
//
