LOCUS       sequence-330390-        6719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 5 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6719)
  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 6719)
  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: pBEL2120
FEATURES             Location/Qualifiers
     source          1..6719
                     /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(3738..3756)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3752..3799
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3844..3862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3930..4034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4035..4103,4104..4895)
                     /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(4253..4272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5066..5654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5555..5574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5808..5825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5840..5979
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5905..5924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6065..6087
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6081..6272)
                     /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 cctgatgttc tgacgctgac
     2581 cctgaccgcg ccggaaagtt acggtattga tgcgggtcag ccgctcattc ttcacggcgt
     2641 gcaggtaggc caggttatcg atcgtaaact caccagcaaa ggcgtcacct ttaccgtcgc
     2701 catcgagcct cagcatcgcg aactggtaaa aggcgatagc aaatttgtcg tcaacagccg
     2761 tgtcgatgtg aaggtggggc tggatggcgt tgagtttctc ggtgccagcg cctcagaatg
     2821 gatcaatggc gggatacgta ttctgccggg cgataaaggt gagatgaaag ccagctatcc
     2881 actgtatgcc aatttggaaa aagcgctgga gaacagcctt agcgatttac ccaccacagg
     2941 cgggcagatt acgcttcacg ctttcgatgc cggaaaactg gcggtcggga tgccaattcg
     3001 ctatctcggt attgatatcg ggcaaatcca gacgctggat ctgattaccg cacgcaatga
     3061 agtacaggca aaggcggtac tttacccgga atatgtccag acctttgctc gcggtggtac
     3121 gcgcttctca gtggtcacac cgcaaatttc ggcagctggc gttgagcatc ttgatactat
     3181 cctccagccg tatatcaacg tcgaaccagg ccggggcaat cctcgccgcg actttgaatt
     3241 acaagaggcc accattactg attcgcgtta cctggatggc ttaagcatta ttgttgaagc
     3301 gccggaagcc ggttcgttag gcatcggtac gcctgtgctg ttccgtggtc tggaagtcgg
     3361 tacggttaca ggaatgacgc tggggacatt gtcagatcgc gtgatgattg cgatgcgcat
     3421 cagtaaacgc tatcaacacc tggtgcgtaa caattccgtg ttctggttgg catcgggtta
     3481 cagtctggac tttggtctga cgggcggcgt agtgaaaacc ggcaccttta accagtttat
     3541 ccgtggcggc atcgccttcg ccacgcctcc gggtacgcca ctggcaccga aagcccagga
     3601 aggcaaacac ttcctgttgc aggaaagtga accgaaagag tggcgtgaat ggggaactgc
     3661 gcttcccaaa taatgcccac ctcgagcaga tccggctgct aacaaagccc gaaaggaagc
     3721 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     3781 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggataattct tgaagacgaa
     3841 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3901 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3961 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4021 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4081 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4141 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4201 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4261 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4321 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4381 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4441 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4501 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4561 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     4621 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4681 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4741 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4801 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4861 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4921 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4981 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5041 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5641 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5701 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5761 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5821 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5881 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5941 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     6001 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6061 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     6121 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     6181 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     6241 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     6301 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     6361 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     6421 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     6481 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     6541 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     6601 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     6661 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggag
//
