LOCUS       sequence-333951-        6713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 2 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6713)
  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 6713)
  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: pBEL2117
FEATURES             Location/Qualifiers
     source          1..6713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      527..1115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1016..1035
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1269..1286
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1301..1440
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1366..1385)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1526..1548
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1542..1733)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2242..2261
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2242..2260
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2292..2314
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5912..5930)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5926..5973
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6018..6036)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6104..6208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6209..6277,6278..6713,1..356)
                     /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(6427..6446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
       61 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
      121 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
      181 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
      241 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
      301 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
      361 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
      421 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      481 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      541 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      601 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      661 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      721 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      781 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      841 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      901 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      961 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1021 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1081 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1141 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1201 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1261 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     1321 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     1381 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     1441 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     1501 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     1561 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     1621 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     1681 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     1741 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     1801 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     1861 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     1921 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     1981 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     2041 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     2101 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     2161 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     2221 gagatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct
     2281 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatggctctt aacacaccac
     2341 aaattacgcc gacaaaaaag ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg
     2401 attaccaacg ttgcccgcaa tgtgacatgc tgtttagcct gcccgagata aattctcatc
     2461 aaagtgccta ttgtccgcgc tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa
     2521 cgcgcctggc ggcaatggcc ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac
     2581 cgctgttgca tatctggctg ttaggcattc gtattgacgc caacgttatg caaggcatct
     2641 ggcaaatgac caaacagggc gatgcgataa cggggtcgat ggtctttttc tgcgttattg
     2701 gtgccccgct cattctggtg acctccatag cttatttatg gtttggtaac cgactgggaa
     2761 tgaatttacg tccggtactg ctgatgcttg agcgacttaa agagtgggta atgctcgata
     2821 tctacctggt cggcattggc gttgcttcta taaaggtaca ggattatgcc catatccagg
     2881 ctggtgtcgg cttgttctct tttgtggcgt tggtgatttt aacgacggtg acgttgtcac
     2941 atcttaatgt cgaagaactg tgggagcgat tttatccgca gcgccccgct acgcgtaggg
     3001 acgagaaact tcgtgtctgc ctagggtgcc attttaccgg ctatccagat cagcgtggtc
     3061 gctgcccgcg ttgccatatc ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct
     3121 gggcggcgct gttagcgtca atcgttttat tgttacctgc caacctgttg cctatttcta
     3181 tcatttatct gaacggagga cggcaggaag atacaattct ttccggaatt atgtcgctgg
     3241 caagtagcaa cattgcggtt gcaggaatcg tgtttatcgc cagtattctg gtcccgttta
     3301 ctaaagtgat cgtcatgttc actttactgt tgagcattca ttttaaatgc cagcaaggtt
     3361 tacgcacacg cattctgtta ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg
     3421 acctgtttgt catatcttta accatgtcgc tgattaatcg cgatcagatc ctcgctttta
     3481 ctatgggacc ggctgcgttt tatttcggcg cagcggtaat tttgactatt cttgctgtgg
     3541 aatggctgga cagccgctta ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg
     3601 actgaagcgc agattaaaaa taaacgccgt atctcacctt tctggctgct gccgttcatc
     3661 gcgctaatga ttgccagttg gctgatttgg gacagttatc aggaccgggg taataccgtc
     3721 accatcgact ttatgtcggc ggatggtatt gttccaggcc gtacgcctgt tcgttatcag
     3781 ggcgttgaag tcggaacagt gcaggatatc agcctcagcg acgatcttcg taagattgaa
     3841 gtcaaggtca gcatcaagtc cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg
     3901 ctggtgacgc caaaagcatc gttggcaggt gtctccgggc tggacgccct cgtcggtggg
     3961 aactatatcg gcatgatgcc gggtaaaggt aaagagcagg atcactttgt cgcactcgat
     4021 acccaaccga aatatcggct ggacaatggc ggcgtaataa taaaactgga actgccgagt
     4081 ggggccggat taaccgccga ctcgacgccg ttaatgtatc aggggctgga agtcggacag
     4141 ctgactaaac tggatttaaa tcctggtggt aaagtcacag gggaaatgac cgttgatccc
     4201 agcgtcgtta ccctgcttcg tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc
     4261 cttagcgatg ctaatctcag cgccctgctg accggcaaaa ccttcgagct ggtgcccggc
     4321 gatggcgagc cacgcaaaga gttcgttgtt gtgccaggcg aaaaagcact gctgcatgaa
     4381 cctgatgttc tgacgctgac cctgaccgcg ccggaaagtt acggtattga tgcgggtcag
     4441 ccgctcattc ttcacggcgt gcaggtaggc caggttatcg atcgtaaact caccagcaaa
     4501 ggcgtcacct ttaccgtcgc catcgagcct cagcatcgcg aactggtaaa aggcgatagc
     4561 aaatttgtcg tcaacagccg tgtcgatgtg aaggtggggc tggatggcgt tgagtttctc
     4621 ggtgccagcg cctcagaatg gatcaatggc gggatacgta ttctgccggg cgataaaggt
     4681 gagatgaaag ccagctatcc actgtatgcc aatttggaaa aagcgctgga gaacagcctt
     4741 agcgatttac ccaccacaac cgtgagtttg agtgcagaga cgctgccgga tgtgcaggca
     4801 ggatcggtag tgctgtaccg taaatttgaa gttggtgaag tgattaccgt gcgtccgcga
     4861 gctaacgcgt ttgatatcga tctgcatatt aagccggagt atcgcaacct tctgaccagc
     4921 aatagcgtgt tctgggcaga aggcggggcg aaagttcagc tgaatggtag tggtctgacc
     4981 gtacaggcat ccccgctctc cagagcatta aagggagcca ttagcttcga caacctcagc
     5041 ggtgccagcg ccagtcagcg taaaggcgat aagcgtattc tgtatgcttc cgaaacagcg
     5101 gcccgtgcgg ttggcgggca gattacgctt cacgctttcg atgccggaaa actggcggtc
     5161 gggatgccaa ttcgctatct cggtattgat atcgggcaaa tccagacgct ggatctgatt
     5221 accgcacgca atgaagtaca ggcaaaggcg gtactttacc cggaatatgt ccagaccttt
     5281 gctcgcggtg gtacgcgctt ctcagtggtc acaccgcaaa tttcggcagc tggcgttgag
     5341 catcttgata ctatcctcca gccgtatatc aacgtcgaac caggccgggg caatcctcgc
     5401 cgcgactttg aattacaaga ggccaccatt actgattcgc gttacctgga tggcttaagc
     5461 attattgttg aagcgccgga agccggttcg ttaggcatcg gtacgcctgt gctgttccgt
     5521 ggtctggaag tcggtacggt tacaggaatg acgctgggga cattgtcaga tcgcgtgatg
     5581 attgcgatgc gcatcagtaa acgctatcaa cacctggtgc gtaacaattc cgtgttctgg
     5641 ttggcatcgg gttacagtct ggactttggt ctgacgggcg gcgtagtgaa aaccggcacc
     5701 tttaaccagt ttatccgtgg cggcatcgcc ttcgccacgc ctccgggtac gccactggca
     5761 ccgaaagccc aggaaggcaa acacttcctg ttgcaggaaa gtgaaccgaa agagtggcgt
     5821 gaatggggaa ctgcgcttcc caaataatgc ccacctcgag cagatccggc tgctaacaaa
     5881 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     5941 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggataa
     6001 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6061 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     6481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atg
//
