LOCUS       sequence-333946-        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     339..358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     592..609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    624..763
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(689..708)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     849..871
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(865..1056)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1565..1584
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1565..1583
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1615..1637
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5241..5259)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5255..5302
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5347..5365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5433..5537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5538..5606,5607..6398)
                     /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(5756..5775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6569..6719,1..438)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
       61 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      541 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      601 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      661 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      721 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
      781 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
      841 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
      901 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
      961 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     1021 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     1081 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     1141 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     1201 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     1261 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     1321 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     1381 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     1441 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     1501 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct cgatcccgcg
     1561 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
     1621 taactttaag aaggagatat acatatggct cttaacacac cacaaattac gccgacaaaa
     1681 aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca acgttgcccg
     1741 caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc ctattgtccg
     1801 cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct ggcggcaatg
     1861 gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt gcatatctgg
     1921 ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat gaccaaacag
     1981 ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc gctcattctg
     2041 gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt acgtccggta
     2101 ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct ggtcggcatt
     2161 ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt cggcttgttc
     2221 tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa tgtcgaagaa
     2281 ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa acttcgtgtc
     2341 tgcctagggt gccattttac cggctatcca gatcagcgtg gtcgctgccc gcgttgccat
     2401 atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc gctgttagcg
     2461 tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta tctgaacgga
     2521 ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag caacattgcg
     2581 gttgcaggaa tcgtgtttat cgccagtatt ctggtcccgt ttactaaagt gatcgtcatg
     2641 ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac acgcattctg
     2701 ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt tgtcatatct
     2761 ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg accggctgcg
     2821 ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct ggacagccgc
     2881 ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag cgcagattaa
     2941 aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa tgattgccag
     3001 ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg actttatgtc
     3061 ggcggatggt attgttccag gccgtacgcc tgttcgttat cagggcgttg aagtcggaac
     3121 agtgcaggat atcagcctca gcgacgatct tcgtaagatt gaagtcaagg tcagcatcaa
     3181 gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga cgccaaaagc
     3241 atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata tcggcatgat
     3301 gccgggtaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac cgaaatatcg
     3361 gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt cgctgaacag
     3421 cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact atgccatcaa
     3481 tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta ccgatctggt
     3541 gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca gtatcagtgg
     3601 cgcgaaggtg aaactggaaa gcctggcggc actggttaac ggtgcgattg ccttcgattc
     3661 accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg aagatctggc
     3721 ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg gattaaccgc
     3781 cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta aactggattt
     3841 aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg ttaccctgct
     3901 tcgtgaaaat acccgcatcg aattacgcaa cccgaaatta tcccttagcg atgctaatct
     3961 cagcgccctg ctgaccggca aaaccttcga gctggtgccc ggcgatggcg agccacgcaa
     4021 agagttcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg ttctgacgct
     4081 gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca ttcttcacgg
     4141 cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca cctttaccgt
     4201 cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg tcgtcaacag
     4261 ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca gcgcctcaga
     4321 atggatcaat ggcgggatac gtattctgcc gggcgataaa ggtgagatga aagccagcta
     4381 tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt tacccaccac
     4441 aggcgggcag attacgcttc acgctttcga tgccggaaaa ctggcggtcg ggatgccaat
     4501 tcgctatctc ggtattgata tcgggcaaat ccagacgctg gatctgatta ccgcacgcaa
     4561 tgaagtacag gcaaaggcgg tactttaccc ggaatatgtc cagacctttg ctcgcggtgg
     4621 tacgcgcttc tcagtggtca caccgcaaat ttcggcagct ggcgttgagc atcttgatac
     4681 tatcctccag ccgtatatca acgtcgaacc aggccggggc aatcctcgcc gcgactttga
     4741 attacaagag gccaccatta ctgattcgcg ttacctggat ggcttaagca ttattgttga
     4801 agcgccggaa gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg gtctggaagt
     4861 cggtacggtt acaggaatga cgctggggac attgtcagat cgcgtgatga ttgcgatgcg
     4921 catcagtaaa cgctatcaac acctggtgcg taacaattcc gtgttctggt tggcatcggg
     4981 ttacagtctg gactttggtc tgacgggcgg cgtagtgaaa accggcacct ttaaccagtt
     5041 tatccgtggc ggcatcgcct tcgccacgcc tccgggtacg ccactggcac cgaaagccca
     5101 ggaaggcaaa cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg aatggggaac
     5161 tgcgcttccc aaataatgcc cacctcgagc agatccggct gctaacaaag cccgaaagga
     5221 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     5281 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataat tcttgaagac
     5341 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5401 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5461 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5521 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5581 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5641 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5701 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5761 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5821 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5881 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5941 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6001 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6061 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6121 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6181 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6241 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6301 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6361 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6421 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6481 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6541 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6601 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6661 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtcctt
//
