LOCUS       sequence-333943-        6680 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid for genetic complementation of LetAB with Delta-Ring 7 LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6680)
  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 6680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pBEL2122
FEATURES             Location/Qualifiers
     source          1..6680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             2964..2986
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3017..3036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3018..3036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3545..3736
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3730..3752)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3838..3977
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3893..3912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3992..4009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4163..4751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4243..4262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4922..5713,5714..5782))
                     /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     5545..5564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5783..5887)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5955..5973
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6018..6065)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6061..6079
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 ggtcagaagg ttgcgatact ccggcttaat atgcagatcg atatcaaacg cgttagctcg
       61 cggacgcacg gtaatcactt caccaacttc aaatttacgg tacagcacta ccgatcctgc
      121 ctgcacatcc ggcagcgtct ctgcactcaa actcacggtt gtggtgggta aatcgctaag
      181 gctgttctcc agcgcttttt ccaaattggc atacagtgga tagctggctt tcatctcacc
      241 tttatcgccc ggcagaatac gtatcccgcc attgatccat tctgaggcgc tggcaccgag
      301 aaactcaacg ccatccagcc ccaccttcac atcgacacgg ctgttgacga caaatttgct
      361 atcgcctttt accagttcgc gatgctgagg ctcgatggcg acggtaaagg tgacgccttt
      421 gctggtgagt ttacgatcga taacctggcc tacctgcacg ccgtgaagaa tgagcggctg
      481 acccgcatca ataccgtaac tttccggcgc ggtcagggtc agcgtcagaa catcaggttc
      541 atgcagcagt gctttttcgc ctggcacaac aacgaactct ttgcgtggct cgccatcgcc
      601 gggcaccagc tcgaaggttt tgccggtcag cagggcgctg agattagcat cgctaaggga
      661 taatttcggg ttgcgtaatt cgatgcgggt attttcacga agcagggtaa cgacgctggg
      721 atcaacggtc atttcccctg tgactttacc accaggattt aaatccagtt tagtcagctg
      781 tccgacttcc agcccctgat acattaacgg cgtcgagtcg gcggttaatc cggccccact
      841 cggcagttcc agttttatta ttacgccacg ctggctgtgg gccagatctt catacagacc
      901 aaaggtatct tccgcctcgg caggtttcga ctcttctggt gaatcgaagg caatcgcacc
      961 gttaaccagt gccgccaggc tttccagttt caccttcgcg ccactgatac tgacgttggc
     1021 atcaacgccg gaaacgttcc agaaacggct accttttttc accagatcgg taaaacgccg
     1081 ctcgatcagg acatcaatca ccacgccttg cttgttggga ttgatggcat agtcgtagac
     1141 ttttcccacc gggatcttgc ggaaatagac caatgaaccg ctgttcagcg aaccgagatc
     1201 gggggcttgc aggtggatca tcagatcgcc attgtccagc cgatatttcg gttgggtatc
     1261 gagtgcgaca aagtgatcct gctctttacc tttacccggc atcatgccga tatagttccc
     1321 accgacgagg gcgtccagcc cggagacacc tgccaacgat gcttttggcg tcaccagcca
     1381 gaactgagtc tcttcgcgca gcgcatcttt catatcggac ttgatgctga ccttgacttc
     1441 aatcttacga agatcgtcgc tgaggctgat atcctgcact gttccgactt caacgccctg
     1501 ataacgaaca ggcgtacggc ctggaacaat accatccgcc gacataaagt cgatggtgac
     1561 ggtattaccc cggtcctgat aactgtccca aatcagccaa ctggcaatca ttagcgcgat
     1621 gaacggcagc agccagaaag gtgagatacg gcgtttattt ttaatctgcg cttcagtcgt
     1681 cgaagcgggc gtttcctgac tcatgtgcat cccaaagtaa gcggctgtcc agccattcca
     1741 cagcaagaat agtcaaaatt accgctgcgc cgaaataaaa cgcagccggt cccatagtaa
     1801 aagcgaggat ctgatcgcga ttaatcagcg acatggttaa agatatgaca aacaggtcga
     1861 gcatcgacca gcgaccaatc caggtcacca tccgcagtaa cagaatgcgt gtgcgtaaac
     1921 cttgctggca tttaaaatga atgctcaaca gtaaagtgaa catgacgatc actttagtaa
     1981 acgggaccag aatactggcg ataaacacga ttcctgcaac cgcaatgttg ctacttgcca
     2041 gcgacataat tccggaaaga attgtatctt cctgccgtcc tccgttcaga taaatgatag
     2101 aaataggcaa caggttggca ggtaacaata aaacgattga cgctaacagc gccgcccagc
     2161 atttttgcag actatgacgg cgacgcaggc gtagcgggat atggcaacgc gggcagcgac
     2221 cacgctgatc tggatagccg gtaaaatggc accctaggca gacacgaagt ttctcgtccc
     2281 tacgcgtagc ggggcgctgc ggataaaatc gctcccacag ttcttcgaca ttaagatgtg
     2341 acaacgtcac cgtcgttaaa atcaccaacg ccacaaaaga gaacaagccg acaccagcct
     2401 ggatatgggc ataatcctgt acctttatag aagcaacgcc aatgccgacc aggtagatat
     2461 cgagcattac ccactcttta agtcgctcaa gcatcagcag taccggacgt aaattcattc
     2521 ccagtcggtt accaaaccat aaataagcta tggaggtcac cagaatgagc ggggcaccaa
     2581 taacgcagaa aaagaccatc gaccccgtta tcgcatcgcc ctgtttggtc atttgccaga
     2641 tgccttgcat aacgttggcg tcaatacgaa tgcctaacag ccagatatgc aacagcggtt
     2701 cgccccaggc aaacggcata agcaacagca tggtgaaggc cattgccgcc aggcgcgtta
     2761 gcgaccagtc gcgcccgtca cgaatttttg cctgacagcg cggacaatag gcactttgat
     2821 gagaatttat ctcgggcagg ctaaacagca tgtcacattg cgggcaacgt tggtaatcac
     2881 cacgcggcag ttcctcgccg attgccctca ctgttatctt ttttgtcggc gtaatttgtg
     2941 gtgtgttaag agccatatgt atatctcctt cttaaagtta aacaaaatta tttctagagg
     3001 gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg agatctcggg
     3061 cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
     3121 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
     3181 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
     3241 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
     3301 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
     3361 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
     3421 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
     3481 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
     3541 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
     3601 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
     3661 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
     3721 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3781 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3841 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3901 gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga
     3961 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4021 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4081 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4561 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4801 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4861 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4921 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4981 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5041 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5101 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5161 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5221 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5281 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5341 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5401 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5461 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5521 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5581 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5641 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5701 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5761 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5821 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5881 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5941 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttatccggat
     6001 atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     6061 gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc tttgttagca
     6121 gccggatctg ctcgaggtgg gcattaatcc aggtaacgcg aatcagtaat ggtggcctct
     6181 tgtaattcaa agtcgcggcg aggattgccc cggcctggtt cgacgttgat atacggctgg
     6241 aggatagtat caagatgctc aacgccagct gccgaaattt gcggtgtgac cactgagaag
     6301 cgcgtaccac cgcgagcaaa ggtctggaca tattccgggt aaagtaccgc ctttgcctgt
     6361 acttcattgc gtgcggtaat cagatccagc gtctggattt gcccgatatc aataccgaga
     6421 tagcgaattg gcatcccgac cgccagtttt ccggcatcga aagcgtgaag cgtaatctgc
     6481 ccgccaaccg cacgggccgc tgtttcggaa gcatacagaa tacgcttatc gcctttacgc
     6541 tgactggcgc tggcaccgct gaggttgtcg aagctaatgg ctccctttaa tgctctggag
     6601 agcggggatg cctgtacggt cagaccacta ccattcagct gaactttcgc cccgccttct
     6661 gcccagaaca cgctattgct
//
