LOCUS       sequence-333948-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 1 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  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 6737)
  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: pBEL2116
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..149
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     65..84
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(164..181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(335..923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(415..434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1094..1885,1886..1954))
                     /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     1717..1736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1955..2059)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2127..2145
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2190..2237)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2233..2251
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             5873..5895
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5926..5945)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5927..5945)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6454..6645
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6639..6661)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
       61 tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca
      121 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
      181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      721 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      961 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1021 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1081 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1141 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1201 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1261 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1321 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1381 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     1441 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1501 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1561 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1621 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1681 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     1741 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1801 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1861 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1921 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1981 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2041 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2101 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     2161 aattatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc
     2221 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg
     2281 gctttgttag cagccggatc tgctcgaggt gggcattatt tgggaagcgc agttccccat
     2341 tcacgccact ctttcggttc actttcctgc aacaggaagt gtttgccttc ctgggctttc
     2401 ggtgccagtg gcgtacccgg aggcgtggcg aaggcgatgc cgccacggat aaactggtta
     2461 aaggtgccgg ttttcactac gccgcccgtc agaccaaagt ccagactgta acccgatgcc
     2521 aaccagaaca cggaattgtt acgcaccagg tgttgatagc gtttactgat gcgcatcgca
     2581 atcatcacgc gatctgacaa tgtccccagc gtcattcctg taaccgtacc gacttccaga
     2641 ccacggaaca gcacaggcgt accgatgcct aacgaaccgg cttccggcgc ttcaacaata
     2701 atgcttaagc catccaggta acgcgaatca gtaatggtgg cctcttgtaa ttcaaagtcg
     2761 cggcgaggat tgccccggcc tggttcgacg ttgatatacg gctggaggat agtatcaaga
     2821 tgctcaacgc cagctgccga aatttgcggt gtgaccactg agaagcgcgt accaccgcga
     2881 gcaaaggtct ggacatattc cgggtaaagt accgcctttg cctgtacttc attgcgtgcg
     2941 gtaatcagat ccagcgtctg gatttgcccg atatcaatac cgagatagcg aattggcatc
     3001 ccgaccgcca gttttccggc atcgaaagcg tgaagcgtaa tctgcccgcc aaccgcacgg
     3061 gccgctgttt cggaagcata cagaatacgc ttatcgcctt tacgctgact ggcgctggca
     3121 ccgctgaggt tgtcgaagct aatggctccc tttaatgctc tggagagcgg ggatgcctgt
     3181 acggtcagac cactaccatt cagctgaact ttcgccccgc cttctgccca gaacacgcta
     3241 ttgctggtca gaaggttgcg atactccggc ttaatatgca gatcgatatc aaacgcgtta
     3301 gctcgcggac gcacggtaat cacttcacca acttcaaatt tacggtacag cactaccgat
     3361 cctgcctgca catccggcag cgtctctgca ctcaaactca cggttgtggt gggtaaatcg
     3421 ctaaggctgt tctccagcgc tttttccaaa ttggcataca gtggatagct ggctttcatc
     3481 tcacctttat cgcccggcag aatacgtatc ccgccattga tccattctga ggcgctggca
     3541 ccgagaaact caacgccatc cagccccacc ttcacatcga cacggctgtt gacgacaaat
     3601 ttgctatcgc cttttaccag ttcgcgatgc tgaggctcga tggcgacggt aaaggtgacg
     3661 cctttgctgg tgagtttacg atcgataacc tggcctacct gcacgccgtg aagaatgagc
     3721 ggctgacccg catcaatacc gtaactttcc ggcgcggtca gggtcagcgt cagaacatca
     3781 ggttcatgca gcagtgcttt ttcgcctggc acaacaacga actctttgcg tggctcgcca
     3841 tcgccgggca ccagctcgaa ggttttgccg gtcagcaggg cgctgagatt agcatcgcta
     3901 agggataatt tcgggttgcg taattcgatg cgggtatttt cacgaagcag ggtaacgacg
     3961 ctgggatcaa cggtcatttc ccctgtgact ttaccaccag gatttaaatc cagtttagtc
     4021 agctgtccga cttccagccc ctgatacatt aacggcgtcg agtcggcggt taatccggcc
     4081 ccactcggca gttccagttt tattattacg ccacgctggc tgtgggccag atcttcatac
     4141 agaccaaagg tatcttccgc ctcggcaggt ttcgactctt ctggtgaatc gaaggcaatc
     4201 gcaccgttaa ccagtgccgc caggctttcc agtttcacct tcgcgccact gatactgacg
     4261 ttggcatcaa cgccggaaac gttccagaaa cggctacctt ttttcaccag atcggtaaaa
     4321 cgccgctcga tcaggacatc aatcaccacg ccttgcttgt tgggattgat ggcatagtcg
     4381 tagacttttc ccaccgggat cttgcggaaa tagaccaatg aaccgctgtt cagcgaaccg
     4441 agatcggggg cttgcaggtg gatcatcaga tcattacccc ggtcctgata actgtcccaa
     4501 atcagccaac tggcaatcat tagcgcgatg aacggcagca gccagaaagg tgagatacgg
     4561 cgtttatttt taatctgcgc ttcagtcgtc gaagcgggcg tttcctgact catgtgcatc
     4621 ccaaagtaag cggctgtcca gccattccac agcaagaata gtcaaaatta ccgctgcgcc
     4681 gaaataaaac gcagccggtc ccatagtaaa agcgaggatc tgatcgcgat taatcagcga
     4741 catggttaaa gatatgacaa acaggtcgag catcgaccag cgaccaatcc aggtcaccat
     4801 ccgcagtaac agaatgcgtg tgcgtaaacc ttgctggcat ttaaaatgaa tgctcaacag
     4861 taaagtgaac atgacgatca ctttagtaaa cggtaccaga atactggcga taaacacgat
     4921 tcctgcaacc gcaatgttgc tacttgccag cgacataatt ccggaaagaa ttgtatcttc
     4981 ctgccgtcct ccgttcagat aaatgataga aataggcaac aggttggcag gtaacaataa
     5041 aacgattgac gctaacagcg ccgcccagca tttttgcaga ctatgacggc gacgcaggcg
     5101 tagcgggata tggcaacgcg ggcagcgacc acgctgatct ggatagccgg taaaatggca
     5161 cccaagacag acacgaagtt tctcgtccct acgcgtagcg gggcgctgcg gataaaatcg
     5221 ctcccacagt tcttcgacat taagatgtga caacgtcacc gtcgttaaaa tcaccaacgc
     5281 cacaaaagag aacaagccga caccagcctg gatatgggca taatcctgta cctttataga
     5341 agcaacgcca atgccgacca ggtagatatc gagcattacc cactctttaa gtcgctcaag
     5401 catcagcagt accggacgta aattcattcc cagtcggtta ccaaaccata aataagctat
     5461 ggaggtcacc agaatgagcg gggcaccaat aacgcagaaa aagaccatcg accccgttat
     5521 cgcatcgccc tgtttggtca tttgccagat gccttgcata acgttggcgt caatacgaat
     5581 gcctaacagc cagatatgca acagcggttc gccccaggca aacggcataa gcaacagcat
     5641 ggtgaaggcc attgccgcca ggcgcgttag cgaccagtcg cgcccgtcac gaatttttgc
     5701 ctgacagcgc ggacaatagg cactttgatg agaatttatc tcgggcaggc taaacagcat
     5761 gtcacattgc gggcaacgtt ggtaatcacc acgcggcagt tcctcgccga ttgccctcac
     5821 tgttatcttt tttgtcggcg taatttgtgg tgtgttaaga gccatatgta tatctccttc
     5881 ttaaagttaa acaaaattat ttctagaggg aaaccgttgt ggtctcccta tagtgagtcg
     5941 tattaatttc gcgggatcga gatctcgggc agcgttgggt cctggccacg ggtgcgcatg
     6001 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     6061 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     6121 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     6181 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     6241 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     6301 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     6361 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     6421 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     6481 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     6541 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     6601 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6661 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6721 tcagcgggtg ttggcgg
//
