LOCUS       sequence-310588-        7043 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Complementation plasmid for LetAB with LetB PLL5 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  AUTHORS   Isom GL, Coudray N, MacRae MR, McManus CT, Ekiert DC, Bhabha G
  TITLE     LetB Structure Reveals a Tunnel for Lipid Transport across the 
            Bacterial Envelope.
  JOURNAL   Cell. 2020 Apr 30;181(3):653-664.e19. doi: 
            10.1016/j.cell.2020.03.030.
  PUBMED    32359438
REFERENCE   2  (bases 1 to 7043)
  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"; doi: 
            "10.1016/j.cell.2020.03"; journalName: "Cell"; date: "2020-04-30- 
            30"; volume: "181"; issue: "3"; pages: "653-664.e19"
COMMENT     Sequence Label: pBEL1890
FEATURES             Location/Qualifiers
     source          1..7043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     316..333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    348..487
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(413..432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     573..595
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(589..780)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1289..1308
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1289..1307
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1339..1361
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5289..5307)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5303..5350
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5395..5413)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5481..5585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5586..5654,5655..6446)
                     /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(5804..5823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6617..7043,1..162)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
       61 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
      361 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
      421 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
      481 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
      541 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
      601 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
      661 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
      721 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
      781 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
      841 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
      901 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
      961 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     1021 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     1081 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     1141 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     1201 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     1261 gctgcccgag atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg
     1321 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat ggctcttaac
     1381 acaccacaaa ttacgccgac aaaaaagata acagtgaggg caatcggcga ggaactgccg
     1441 cgtggtgatt accaacgttg cccgcaatgt gacatgctgt ttagcctgcc cgagataaat
     1501 tctcatcaaa gtgcctattg tccgcgctgt caggcaaaaa ttcgtgacgg gcgcgactgg
     1561 tcgctaacgc gcctggcggc aatggccttc accatgctgt tgcttatgcc gtttgcctgg
     1621 ggcgaaccgc tgttgcatat ctggctgtta ggcattcgta ttgacgccaa cgttatgcaa
     1681 ggcatctggc aaatgaccaa acagggcgat gcgataacgg ggtcgatggt ctttttctgc
     1741 gttattggtg ccccgctcat tctggtgacc tccatagctt atttatggtt tggtaaccga
     1801 ctgggaatga atttacgtcc ggtactgctg atgcttgagc gacttaaaga gtgggtaatg
     1861 ctcgatatct acctggtcgg cattggcgtt gcttctataa aggtacagga ttatgcccat
     1921 atccaggctg gtgtcggctt gttctctttt gtggcgttgg tgattttaac gacggtgacg
     1981 ttgtcacatc ttaatgtcga agaactgtgg gagcgatttt atccgcagcg ccccgctacg
     2041 cgtagggacg agaaacttcg tgtctgccta gggtgccatt ttaccggcta tccagatcag
     2101 cgtggtcgct gcccgcgttg ccatatcccg ctacgcctgc gtcgccgtca tagtctgcaa
     2161 aaatgctggg cggcgctgtt agcgtcaatc gttttattgt tacctgccaa cctgttgcct
     2221 atttctatca tttatctgaa cggaggacgg caggaagata caattctttc cggaattatg
     2281 tcgctggcaa gtagcaacat tgcggttgca ggaatcgtgt ttatcgccag tattctggtc
     2341 ccgtttacta aagtgatcgt catgttcact ttactgttga gcattcattt taaatgccag
     2401 caaggtttac gcacacgcat tctgttactg cggatggtga cctggattgg tcgctggtcg
     2461 atgctcgacc tgtttgtcat atctttaacc atgtcgctga ttaatcgcga tcagatcctc
     2521 gcttttacta tgggaccggc tgcgttttat ttcggcgcag cggtaatttt gactattctt
     2581 gctgtggaat ggctggacag ccgcttactt tgggatgcac atgagtcagg aaacgcccgc
     2641 ttcgacgact gaagcgcaga ttaaaaataa acgccgtatc tcacctttct ggctgctgcc
     2701 gttcatcgcg ctaatgattg ccagttggct gatttgggac agttatcagg accggggtaa
     2761 taccgtcacc atcgacttta tgtcggcgga tggtattgtt ccaggccgta cgcctgttcg
     2821 ttatcagggc gttgaagtcg gaacagtgca ggacatcagc ctcagcgacg atcttcgtaa
     2881 gattgaagtc aaggtcagca tcaagtccga tatgaaagat gcgctgcgcg aagagactca
     2941 gttctggctg gtgacgccaa aagcatcgtt ggcaggtgtc tccgggctgg acgccctcgt
     3001 cggtgggaac tatatcggca tgatgcccgg gaaaggtaaa gagcaggatc actttgtcgc
     3061 actcgatacc caaccgaaat atcggctgga caatggcgat ctgatgatcc acctgcaagc
     3121 ccccgatctc ggttcgctga acagcggttc attggtctat ttccgcaaga tcccggtggg
     3181 aaaagtctac gactatgcca tcaatcccaa caagcaaggc gtggtgattg atgtcctgat
     3241 cgagcggcgt tttaccgatc tggtgaaaaa aggtagccgt ttctggaacg tttccggcgt
     3301 tgatgccaac gtcagtatca gtggcgcgaa ggtgaaactg gaaagcttgg cggcactggt
     3361 taacggtgcg attgccttcg attcaccaga agagtcgaaa cctgccgagg cggaagatac
     3421 ctttggtctg tatgaagatc tggcccacag ccagcgtggc gtaataataa aactggaact
     3481 gccgagtggg gccggattaa ccgccgactc gacgccgtta atgtatcagg ggctggaagt
     3541 cggacagctg actaaactgg atttaaatcc tggtggtaaa gtcacagggg aaatgaccgt
     3601 tgatcccagc gtcgttaccc tgcttcgtga aaatacccgc atcgaattac gcaacccgaa
     3661 attatccctt agcgatgcta atctcagcgc cctgctgacc ggcaaaacct tcgagctggt
     3721 gcccggcgat ggcgagccac gcaaagaatt cgttgttgtg ccaggcgaaa aagcactgct
     3781 gcatgaacct gatgttctga cgctgaccct gaccgcgccg gaaagttacg gtattgatgc
     3841 gggtcagccg ctcattcttc acggcgtgca ggtaggccag gttatcgatc gtaaactcac
     3901 cagcaaaggc gtcaccttta ccgtcgccat cgagcctcag catcgcgaac tggtaaaagg
     3961 cgatagcaaa tttgtcgtca acagccgtgt cgatgtgaag gtggggctgg atggcgttga
     4021 gtttctcggt gccagcgcct cagaatggat caatggcggg atccgtattc tgccgggcga
     4081 taaaggtgag atgaaagcca gctatccact gtatgccaat ttggaaaaag cgctggagaa
     4141 cagccttagc gatttaccca ccacaaccgt gagtttgagt gcagagacgc tgccggatgt
     4201 gcaggcagga tcggtagtgc tgtaccgtaa atttgaagtt ggtgaagtga ttaccgtgcg
     4261 tccgcgagct aacgcgtttg acatcgatct gcatattaag ccggagtatc gcaaccttct
     4321 gaccagcaat agcgtgttct gggcagaagg cggggcggga ggggcatccc cgctctccag
     4381 agctctaaag ggagccatta gcttcgacaa cctcagcggt gccagcgcca gtcagcgtaa
     4441 aggcgataag cgtattctgt atgcttccga aacagcggcc cgtgcggttg gcgggcagat
     4501 tacgcttcac gctttcgatg ccggaaaact ggcggtcggg atgccaattc gctatctcgg
     4561 tattgacatc gggcaaatcc agacgctgga tctgattacc gcacgcaatg aagtacaggc
     4621 aaaggcggta ctttacccgg aatatgtcca gacctttgct cgcggtggta cgcgcttctc
     4681 agtggtcaca ccgcaaattt cggcagctgg cgttgagcat cttgatacta tcctccagcc
     4741 gtatatcaac gtcgaaccag gccggggcaa tcctcgccgc gactttgaat tacaagaggc
     4801 caccattact gattcgcggt acctggatgg cttaagcatt attgttgaag cgccggaagc
     4861 cggttcgtta ggcatcggta cgcctgtgct gttccgtggt ctggaagtcg gtacggttac
     4921 aggaatgacg ctggggacat tgtcagatcg cgtgatgatt gcgatgcgca tcagtaaacg
     4981 ctatcaacac ctggtgcgta acaattccgt cttctggttg gcatcgggtt acagtctgga
     5041 ctttggtctg acgggcggcg tagtgaaaac cggcaccttt aaccagttta tccgtggcgg
     5101 catcgccttc gccacgcctc cgggtacgcc actggcaccg aaagcccagg aaggcaaaca
     5161 cttcctgttg caggaaagtg aaccgaaaga gtggcgtgaa tggggaactg cgcttcccaa
     5221 ataatgccca cctcgagcag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     5281 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     5341 gggttttttg ctgaaaggag gaactatatc cggataattc ttgaagacga aagggcctcg
     5401 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5701 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5761 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5821 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5881 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5941 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6001 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6061 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6121 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6181 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6241 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6301 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6361 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6421 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6481 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7021 ccacgcttcc cgaagggaga aag
//
