LOCUS       sequence-333944-        6734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 6 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6734)
  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 6734)
  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: pBEL2121
FEATURES             Location/Qualifiers
     source          1..6734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     243..260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    275..414
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(340..359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     500..522
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(516..707)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1216..1235
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1216..1234
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1266..1288
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4907..4925)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4921..4968
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5013..5031)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5099..5203
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5204..5272,5273..6064)
                     /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(5422..5441)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6235..6734,1..89)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(6724..6734,1..9)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
       61 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      241 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
      301 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
      361 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
      421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      541 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
      601 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
      661 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
      721 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
      781 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
      841 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
      901 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
      961 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     1021 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     1081 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     1141 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatc
     1201 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
     1261 ataattttgt ttaactttaa gaaggagata tacatatggc tcttaacaca ccacaaatta
     1321 cgccgacaaa aaagataaca gtgagggcaa tcggcgagga actgccgcgt ggtgattacc
     1381 aacgttgccc gcaatgtgac atgctgttta gcctgcccga gataaattct catcaaagtg
     1441 cctattgtcc gcgctgtcag gcaaaaattc gtgacgggcg cgactggtcg ctaacgcgcc
     1501 tggcggcaat ggccttcacc atgctgttgc ttatgccgtt tgcctggggc gaaccgctgt
     1561 tgcatatctg gctgttaggc attcgtattg acgccaacgt tatgcaaggc atctggcaaa
     1621 tgaccaaaca gggcgatgcg ataacggggt cgatggtctt tttctgcgtt attggtgccc
     1681 cgctcattct ggtgacctcc atagcttatt tatggtttgg taaccgactg ggaatgaatt
     1741 tacgtccggt actgctgatg cttgagcgac ttaaagagtg ggtaatgctc gatatctacc
     1801 tggtcggcat tggcgttgct tctataaagg tacaggatta tgcccatatc caggctggtg
     1861 tcggcttgtt ctcttttgtg gcgttggtga ttttaacgac ggtgacgttg tcacatctta
     1921 atgtcgaaga actgtgggag cgattttatc cgcagcgccc cgctacgcgt agggacgaga
     1981 aacttcgtgt ctgcctaggg tgccatttta ccggctatcc agatcagcgt ggtcgctgcc
     2041 cgcgttgcca tatcccgcta cgcctgcgtc gccgtcatag tctgcaaaaa tgctgggcgg
     2101 cgctgttagc gtcaatcgtt ttattgttac ctgccaacct gttgcctatt tctatcattt
     2161 atctgaacgg aggacggcag gaagatacaa ttctttccgg aattatgtcg ctggcaagta
     2221 gcaacattgc ggttgcagga atcgtgttta tcgccagtat tctggtcccg tttactaaag
     2281 tgatcgtcat gttcacttta ctgttgagca ttcattttaa atgccagcaa ggtttacgca
     2341 cacgcattct gttactgcgg atggtgacct ggattggtcg ctggtcgatg ctcgacctgt
     2401 ttgtcatatc tttaaccatg tcgctgatta atcgcgatca gatcctcgct tttactatgg
     2461 gaccggctgc gttttatttc ggcgcagcgg taattttgac tattcttgct gtggaatggc
     2521 tggacagccg cttactttgg gatgcacatg agtcaggaaa cgcccgcttc gacgactgaa
     2581 gcgcagatta aaaataaacg ccgtatctca cctttctggc tgctgccgtt catcgcgcta
     2641 atgattgcca gttggctgat ttgggacagt tatcaggacc ggggtaatac cgtcaccatc
     2701 gactttatgt cggcggatgg tattgttcca ggccgtacgc ctgttcgtta tcagggcgtt
     2761 gaagtcggaa cagtgcagga tatcagcctc agcgacgatc ttcgtaagat tgaagtcaag
     2821 gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag agactcagtt ctggctggtg
     2881 acgccaaaag catcgttggc aggtgtctcc gggctggacg ccctcgtcgg tgggaactat
     2941 atcggcatga tgccgggtaa aggtaaagag caggatcact ttgtcgcact cgatacccaa
     3001 ccgaaatatc ggctggacaa tggcgatctg atgatccacc tgcaagcccc cgatctcggt
     3061 tcgctgaaca gcggttcatt ggtctatttc cgcaagatcc cggtgggaaa agtctacgac
     3121 tatgccatca atcccaacaa gcaaggcgtg gtgattgatg tcctgatcga gcggcgtttt
     3181 accgatctgg tgaaaaaagg tagccgtttc tggaacgttt ccggcgttga tgccaacgtc
     3241 agtatcagtg gcgcgaaggt gaaactggaa agcctggcgg cactggttaa cggtgcgatt
     3301 gccttcgatt caccagaaga gtcgaaacct gccgaggcgg aagatacctt tggtctgtat
     3361 gaagatctgg cccacagcca gcgtggcgta ataataaaac tggaactgcc gagtggggcc
     3421 ggattaaccg ccgactcgac gccgttaatg tatcaggggc tggaagtcgg acagctgact
     3481 aaactggatt taaatcctgg tggtaaagtc acaggggaaa tgaccgttga tcccagcgtc
     3541 gttaccctgc ttcgtgaaaa tacccgcatc gaattacgca acccgaaatt atcccttagc
     3601 gatgctaatc tcagcgccct gctgaccggc aaaaccttcg agctggtgcc cggcgatggc
     3661 gagccacgca aagagttcgt tgttgtgcca ggcgaaaaag cactgctgca tgaacctgat
     3721 gttctgacgc tgaccctgac cgcgccggaa agttacggta ttgatgcggg tcagccgctc
     3781 attcttcacg gcgtgcaggt aggccaggtt atcgatcgta aactcaccag caaaggcgtc
     3841 acctttaccg tcgccatcga gcctcagcat cgcgaactgg taaaaggcga tagcaaattt
     3901 gtcgtcaaca gccgtgtcga tgtgaaggtg gggctggatg gcgttgagtt tctcggtgcc
     3961 agcgcctcag aatggatcaa tggcgggata cgtattctgc cgggcgataa aggtgagatg
     4021 aaagccagct atccactgta tgccaatttg gaaaaagcgc tggagaacag ccttagcgat
     4081 ttacccacca caaccgtgag tttgagtgca gagacgctgc cggatgtgca ggcaggatcg
     4141 gtagtgctgt accgtaaatt tgaagttggt gaagtgatta ccgtgcgtcc gcgagctaac
     4201 gcgtttgata tcgatctgca tattaagccg gagtatcgca accttctgac cagcaatagc
     4261 gtgttctggg cagaaggcgg ggcgaaagtt cagctgaatg gtagtggtct gaccgtacag
     4321 gcatccccgc tctccagagc attaaaggga gccattagct tcgacaacct cagcggtgcc
     4381 agcgccagtc agcgtaaagg cgataagcgt attctgtatg cttccgaaac agcggcccgt
     4441 gcggttggct taagcattat tgttgaagcg ccggaagccg gttcgttagg catcggtacg
     4501 cctgtgctgt tccgtggtct ggaagtcggt acggttacag gaatgacgct ggggacattg
     4561 tcagatcgcg tgatgattgc gatgcgcatc agtaaacgct atcaacacct ggtgcgtaac
     4621 aattccgtgt tctggttggc atcgggttac agtctggact ttggtctgac gggcggcgta
     4681 gtgaaaaccg gcacctttaa ccagtttatc cgtggcggca tcgccttcgc cacgcctccg
     4741 ggtacgccac tggcaccgaa agcccaggaa ggcaaacact tcctgttgca ggaaagtgaa
     4801 ccgaaagagt ggcgtgaatg gggaactgcg cttcccaaat aatgcccacc tcgagcagat
     4861 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     4921 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     4981 actatatccg gataattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     5041 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5101 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5161 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5221 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5281 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5341 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5401 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     5461 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5521 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5581 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5641 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5701 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     5761 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5821 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5881 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5941 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6001 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6061 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6121 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6181 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6241 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6301 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6361 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6421 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6481 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6541 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6601 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6661 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6721 agggggaaac gcct
//
