LOCUS       sequence-345261-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetAB with 6PLL3-hairpin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7070)
  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 7070)
  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: pBEL2412
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..54)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      848..1436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1337..1356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1590..1607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1622..1761
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1687..1706)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1847..1869
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1863..2054)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2563..2582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2563..2581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2613..2635
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6590..6608)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6604..6651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6696..6714)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6782..6886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6887..6955,6956..7070,1..677)
                     /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 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
       61 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      361 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      781 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      961 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     1021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1561 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1621 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     1681 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     1741 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     1801 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     1861 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     1921 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     1981 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     2041 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     2101 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     2161 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     2221 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     2281 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     2341 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     2401 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     2461 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt
     2521 ggccaggacc caacgctgcc cgagatctcg atcccgcgaa attaatacga ctcactatag
     2581 ggagaccaca acggtttccc tctagaaata attttgttta actttaagaa ggagatatac
     2641 atatggctct taacacacca caaattacgc cgacaaaaaa gataacagtg agggcaatcg
     2701 gcgaggaact gccgcgtggt gattaccaac gttgcccgca atgtgacatg ctgtttagcc
     2761 tgcccgagat aaattctcat caaagtgcct attgtccgcg ctgtcaggca aaaattcgtg
     2821 acgggcgcga ctggtcgcta acgcgcctgg cggcaatggc cttcaccatg ctgttgctta
     2881 tgccgtttgc ctggggcgaa ccgctgttgc atatctggct gttaggcatt cgtattgacg
     2941 ccaacgttat gcaaggcatc tggcaaatga ccaaacaggg cgatgcgata acggggtcga
     3001 tggtcttttt ctgcgttatt ggtgccccgc tcattctggt gacctccata gcttatttat
     3061 ggtttggtaa ccgactggga atgaatttac gtccggtact gctgatgctt gagcgactta
     3121 aagagtgggt aatgctcgat atctacctgg tcggcattgg cgttgcttct ataaaggtac
     3181 aggattatgc ccatatccag gctggtgtcg gcttgttctc ttttgtggcg ttggtgattt
     3241 taacgacggt gacgttgtca catcttaatg tcgaagaact gtgggagcga ttttatccgc
     3301 agcgccccgc tacgcgtagg gacgagaaac ttcgtgtctg tcttgggtgc cattttaccg
     3361 gctatccaga tcagcgtggt cgctgcccgc gttgccatat cccgctacgc ctgcgtcgcc
     3421 gtcatagtct gcaaaaatgc tgggcggcgc tgttagcgtc aatcgtttta ttgttacctg
     3481 ccaacctgtt gcctatttct atcatttatc tgaacggagg acggcaggaa gatacaattc
     3541 tttccggaat tatgtcgctg gcaagtagca acattgcggt tgcaggaatc gtgtttatcg
     3601 ccagtattct ggtaccgttt actaaagtga tcgtcatgtt cactttactg ttgagcattc
     3661 attttaaatg ccagcaaggt ttacgcacac gcattctgtt actgcggatg gtgacctgga
     3721 ttggtcgctg gtcgatgctc gacctgtttg tcatatcttt aaccatgtcg ctgattaatc
     3781 gcgatcagat cctcgctttt actatgggac cggctgcgtt ttatttcggc gcagcggtaa
     3841 ttttgactat tcttgctgtg gaatggctgg acagccgctt actttgggat gcacatgagt
     3901 caggaaacgc ccgcttcgac gactgaagcg cagattaaaa ataaacgccg tatctcacct
     3961 ttctggctgc tgccgttcat cgcgctaatg attgccagtt ggctgatttg ggacagttat
     4021 caggaccggg gtaataccgt caccatcgac tttatgtcgg cggatggtat tgttccaggc
     4081 cgtacgcctg ttcgttatca gggcgttgaa gtcggaacag tgcaggatat cagcctcagc
     4141 gacgatcttc gtaagattga agtcaaggtc agcatcaagt ccgatatgaa agatgcgctg
     4201 cgcgaagaga ctcagttctg gctggtgacg ccaaaagcat cgttggcagg tgtctccggg
     4261 ctggacgccc tcgtcggtgg gaactatatc ggcatgatgc cgggtaaagg taaagagcag
     4321 gatcactttg tcgcactcga tacccaaccg aaatatcggc tggacaatgg cgatctgatg
     4381 atccacctgc aagcccccga tctcggttcg ctgaacagcg gttcattggt ctatttccgc
     4441 aagatcccgg tgggaaaagt ctacgactat gccatcaatc ccaacaagca aggcgtggtg
     4501 attgatgtcc tgatcgagcg gcgttttacc gatctggtga aaaaaggtag ccgtttctgg
     4561 aacgtttccg gcgttgatgc caacgtcagt atcagtggcg cgaaggtgaa actggaaagc
     4621 ctggcggcac tggttaacgg tgcgattgcc ttcgattcac cagaagagtc gaaacctgcc
     4681 gaggcggaag atacctttgg tctgtatgaa gatctggccc acagccagcg tggcgtaata
     4741 ataaaactgg aactgccgag tggggccgga ttaaccgccg actcgacgcc gttaatgtat
     4801 caggggctgg aagtcggaca gctgactaaa ctggatttaa atcctggtgg taaagtcaca
     4861 ggggaaatga ccgttgatcc cagcgtcgtt accctgcttc gtgaaaatac ccgcatcgaa
     4921 ttacgcaacc cgaaattatc ccttagcgat gctaatctca gcgccctgct gaccggcaaa
     4981 accttcgagc tggtgcccgg cgatggcgag ccacgcaaag agttcgttgt tgtgccaggc
     5041 gaaaaagcac tgctgcatga acctgatgtt ctgacgctga ccctgaccgc gccggaaagt
     5101 tacggtattg atgcgggtca gccgctcatt cttcacggcg tgcaggtagg ccaggttatc
     5161 gatcgtaaac tcaccagcaa aggcgtcacc tttaccgtcg ccatcgagcc tcagcatcgc
     5221 gaactggtaa aaggcgatag caaatttgtc gtcaacagcc gtgtcgatgt gaaggtgggg
     5281 ctggatggcg ttgagtttct cggtgccagc gcctcagaat ggatcaatgg cgggatacgt
     5341 attctgccgg gcgataaagg tgagatgaaa gccagctatc cactgtatgc caatttggaa
     5401 aaagcgctgg agaacagcct tagcgattta cccaccacaa ccgtgagttt gagtgcagag
     5461 acgctgccgg atgtgcaggc aggatcggta gtgctgtacc gtaaatttga agttggtgaa
     5521 gtgattaccg tgcgtccgcg agctaacgcg tttgatatcg atctgcatat taagccggag
     5581 tatcgcaacc ttctgaccag caatagcgtg ttctgggcag aaggcggggc gaaagttcag
     5641 ctgaatggta gtggtctgac cgtacaggca tccccgctct ccagagcatt aaagggagcc
     5701 attagcttcg acaacctcag cggtgccagc gccagtcagc gtaaaggcga taagcgtatt
     5761 ctgtatgctt ccgaaacagc ggcccgtgcg gttggcgggc agattacgct tcacgctttc
     5821 gatgccggaa aactggcggt cgggatgcca attcgctatc tcggtattga tatcgggcaa
     5881 atccagacgc tggatctgat taccgcacgc aatgaagtac aggcaaaggc ggtactttac
     5941 ccggaatatg tccagacctt tgctcgcggt ggtacgcgct tctcagtggt cacaccgaaa
     6001 ttatccctta gcgatgctaa tcttgatact atcctccagc cgtatatcaa cgtcgaacca
     6061 ggccggggca atcctcgccg cgactttgaa ttacaagagg ccaccattac tgattcgcgt
     6121 tacctggatg gcttaagcat tattgttgaa gcgccggaag ccggttcgtt aggcatcggt
     6181 acgcctgtgc tgttccgtgg tctggaagtc ggtacggtta caggaatgac gctggggaca
     6241 ttgtcagatc gcgtgatgat tgcgatgcgc atcagtaaac gctatcaaca cctggtgcgt
     6301 aacaattccg tcttctggtt ggcatcgggt tacagtctgg actttggtct gacgggcggc
     6361 gtagtgaaaa ccggcacctt taaccagttt atccgtggcg gcatcgcctt cgccacgcct
     6421 ccgggtacgc cactggcacc gaaagcccag gaaggcaaac acttcctgtt gcaggaaagt
     6481 gaaccgaaag agtggcgtga atggggaact gcgcttccca aataatgccc acctcgagca
     6541 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     6601 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     6661 ggaactatat ccggataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     6721 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6781 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6841 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6901 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6961 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7021 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
//
