LOCUS       sequence-330389-        6701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 4 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  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 6701)
  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: pBEL2119
FEATURES             Location/Qualifiers
     source          1..6701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3720..3738)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3734..3781
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3826..3844)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3912..4016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4017..4085,4086..4877)
                     /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(4235..4254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5048..5636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5537..5556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5790..5807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5822..5961
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5887..5906)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6047..6069
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6063..6254)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatggctctt aacacaccac aaattacgcc gacaaaaaag
      181 ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa
      241 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      301 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      361 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      421 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      481 gatgcgataa cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg
      541 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      601 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      661 gttgcttcta taaaggtaca ggattatgcc catatccagg ctggtgtcgg cttgttctct
      721 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
      781 tgggagcgat tttatccgca gcgccccgct acgcgtaggg acgagaaact tcgtgtctgc
      841 ctagggtgcc attttaccgg ctatccagat cagcgtggtc gctgcccgcg ttgccatatc
      901 ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct gggcggcgct gttagcgtca
      961 atcgttttat tgttacctgc caacctgttg cctatttcta tcatttatct gaacggagga
     1021 cggcaggaag atacaattct ttccggaatt atgtcgctgg caagtagcaa cattgcggtt
     1081 gcaggaatcg tgtttatcgc cagtattctg gtcccgttta ctaaagtgat cgtcatgttc
     1141 actttactgt tgagcattca ttttaaatgc cagcaaggtt tacgcacacg cattctgtta
     1201 ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg acctgtttgt catatcttta
     1261 accatgtcgc tgattaatcg cgatcagatc ctcgctttta ctatgggacc ggctgcgttt
     1321 tatttcggcg cagcggtaat tttgactatt cttgctgtgg aatggctgga cagccgctta
     1381 ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa
     1441 taaacgccgt atctcacctt tctggctgct gccgttcatc gcgctaatga ttgccagttg
     1501 gctgatttgg gacagttatc aggaccgggg taataccgtc accatcgact ttatgtcggc
     1561 ggatggtatt gttccaggcc gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt
     1621 gcaggatatc agcctcagcg acgatcttcg taagattgaa gtcaaggtca gcatcaagtc
     1681 cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg ctggtgacgc caaaagcatc
     1741 gttggcaggt gtctccgggc tggacgccct cgtcggtggg aactatatcg gcatgatgcc
     1801 gggtaaaggt aaagagcagg atcactttgt cgcactcgat acccaaccga aatatcggct
     1861 ggacaatggc gatctgatga tccacctgca agcccccgat ctcggttcgc tgaacagcgg
     1921 ttcattggtc tatttccgca agatcccggt gggaaaagtc tacgactatg ccatcaatcc
     1981 caacaagcaa ggcgtggtga ttgatgtcct gatcgagcgg cgttttaccg atctggtgaa
     2041 aaaaggtagc cgtttctgga acgtttccgg cgttgatgcc aacgtcagta tcagtggcgc
     2101 gaaggtgaaa ctggaaagcc tggcggcact ggttaacggt gcgattgcct tcgattcacc
     2161 agaagagtcg aaacctgccg aggcggaaga tacctttggt ctgtatgaag atctggccca
     2221 cagccagcgt ggcgtaataa taaaactgga actgccgagt ggggccggat taaccgccga
     2281 ctcgacgccg ttaatgtatc aggggctgga agtcggacag ctgactaaac tggatttaaa
     2341 tcctggtggt aaagtcacag gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg
     2401 tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc cttagcgatg ctaatctcag
     2461 cgccctgctg accggcaaaa ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga
     2521 gttcgttgtt gtgccaggcg aaaaagcact gctgcatgaa cctgataccg tgagtttgag
     2581 tgcagagacg ctgccggatg tgcaggcagg atcggtagtg ctgtaccgta aatttgaagt
     2641 tggtgaagtg attaccgtgc gtccgcgagc taacgcgttt gatatcgatc tgcatattaa
     2701 gccggagtat cgcaaccttc tgaccagcaa tagcgtgttc tgggcagaag gcggggcgaa
     2761 agttcagctg aatggtagtg gtctgaccgt acaggcatcc ccgctctcca gagcattaaa
     2821 gggagccatt agcttcgaca acctcagcgg tgccagcgcc agtcagcgta aaggcgataa
     2881 gcgtattctg tatgcttccg aaacagcggc ccgtgcggtt ggcgggcaga ttacgcttca
     2941 cgctttcgat gccggaaaac tggcggtcgg gatgccaatt cgctatctcg gtattgatat
     3001 cgggcaaatc cagacgctgg atctgattac cgcacgcaat gaagtacagg caaaggcggt
     3061 actttacccg gaatatgtcc agacctttgc tcgcggtggt acgcgcttct cagtggtcac
     3121 accgcaaatt tcggcagctg gcgttgagca tcttgatact atcctccagc cgtatatcaa
     3181 cgtcgaacca ggccggggca atcctcgccg cgactttgaa ttacaagagg ccaccattac
     3241 tgattcgcgt tacctggatg gcttaagcat tattgttgaa gcgccggaag ccggttcgtt
     3301 aggcatcggt acgcctgtgc tgttccgtgg tctggaagtc ggtacggtta caggaatgac
     3361 gctggggaca ttgtcagatc gcgtgatgat tgcgatgcgc atcagtaaac gctatcaaca
     3421 cctggtgcgt aacaattccg tgttctggtt ggcatcgggt tacagtctgg actttggtct
     3481 gacgggcggc gtagtgaaaa ccggcacctt taaccagttt atccgtggcg gcatcgcctt
     3541 cgccacgcct ccgggtacgc cactggcacc gaaagcccag gaaggcaaac acttcctgtt
     3601 gcaggaaagt gaaccgaaag agtggcgtga atggggaact gcgcttccca aataatgccc
     3661 acctcgagca gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     3721 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     3781 gctgaaagga ggaactatat ccggataatt cttgaagacg aaagggcctc gtgatacgcc
     3841 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3961 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     4261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4561 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5161 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5701 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5761 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5821 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac
     5881 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     5941 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6001 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6061 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     6121 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     6181 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     6241 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     6301 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     6361 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     6421 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     6481 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     6541 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     6601 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     6661 aggcaacccc gccagcctag ccgggtcctc aacgacagga g
//
