LOCUS       sequence-270803-        6020 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Complementation plasmid for LetAB with 4 ring (rings 1347) LetB 
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6020)
  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 6020)
  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: pBEL2042
FEATURES             Location/Qualifiers
     source          1..6020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3039..3057)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3053..3100
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3145..3163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3231..3335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3336..3404,3405..4196)
                     /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(3554..3573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4367..4955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4856..4875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5109..5126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5141..5280
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5206..5225)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5366..5388
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5382..5573)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatggctctt aacacaccac aaattacgcc gacaaaaaag
      181 ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa
      241 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      301 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      361 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      421 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      481 gatgcgataa cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg
      541 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      601 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      661 gttgcttcta taaaggtaca ggattatgcc catatccagg ctggtgtcgg cttgttctct
      721 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
      781 tgggagcgat tttatccgca gcgccccgct acgcgtaggg acgagaaact tcgtgtctgc
      841 ctagggtgcc attttaccgg ctatccagat cagcgtggtc gctgcccgcg ttgccatatc
      901 ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct gggcggcgct gttagcgtca
      961 atcgttttat tgttacctgc caacctgttg cctatttcta tcatttatct gaacggagga
     1021 cggcaggaag atacaattct ttccggaatt atgtcgctgg caagtagcaa cattgcggtt
     1081 gcaggaatcg tgtttatcgc cagtattctg gtcccgttta ctaaagtgat cgtcatgttc
     1141 actttactgt tgagcattca ttttaaatgc cagcaaggtt tacgcacacg cattctgtta
     1201 ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg acctgtttgt catatcttta
     1261 accatgtcgc tgattaatcg cgatcagatc ctcgctttta ctatgggacc ggctgcgttt
     1321 tatttcggcg cagcggtaat tttgactatt cttgctgtgg aatggctgga cagccgctta
     1381 ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa
     1441 taaacgccgt atctcacctt tctggctgct gccgttcatc gcgctaatga ttgccagttg
     1501 gctgatttgg gacagttatc aggaccgggg taataccgtc accatcgact ttatgtcggc
     1561 ggatggtatt gttccaggcc gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt
     1621 gcaggacatc agcctcagcg acgatcttcg taagattgaa gtcaaggtca gcatcaagtc
     1681 cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg ctggtgacgc caaaagcatc
     1741 gttggcaggt gtctccgggc tggacgccct cgtcggtggg aactatatcg gcatgatgcc
     1801 cgggaaaggt aaagagcagg atcactttgt cgcactcgat acccaaccga aatatcggct
     1861 ggacaatggc ggcgtaataa taaaactgga actgccgagt ggggccggat taaccgccga
     1921 ctcgacgccg ttaatgtatc aggggctgga agtcggacag ctgactaaac tggatttaaa
     1981 tcctggtggt aaagtcacag gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg
     2041 tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc cttagcgatg ctaatctcag
     2101 cgccctgctg accggcaaaa ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga
     2161 attcgttgtt gtgccaggcg aaaaagcact gctgcatgaa cctgatgttc tgacgctgac
     2221 cctgaccgcg ccggaaagtt acggtattga tgcgggtcag ccgctcattc ttcacggcgt
     2281 gcaggtaggc caggttatcg atcgtaaact caccagcaaa ggcgtcacct ttaccgtcgc
     2341 catcgagcct cagcatcgcg aactggtaaa aggcgatagc aaatttgtcg tcaacagccg
     2401 tgtcgatgtg aaggtggggc tggatggcgt tgagtttctc ggtgccagcg cctcagaatg
     2461 gatcaatggc gggatccgta ttctgccggg cgataaaggt gagatgaaag ccagctatcc
     2521 actgtatgcc aatttggaaa aagcgctgga gaacagcctt agcgatttac ccaccacagg
     2581 cttaagcatt attgttgaag cgccggaagc cggttcgtta ggcatcggta cgcctgtgct
     2641 gttccgtggt ctggaagtcg gtacggttac aggaatgacg ctggggacat tgtcagatcg
     2701 cgtgatgatt gcgatgcgca tcagtaaacg ctatcaacac ctggtgcgta acaattccgt
     2761 cttctggttg gcatcgggtt acagtctgga ctttggtctg acgggcggcg tagtgaaaac
     2821 cggcaccttt aaccagttta tccgtggcgg catcgccttc gccacgcctc cgggtacgcc
     2881 actggcaccg aaagcccagg aaggcaaaca cttcctgttg caggaaagtg aaccgaaaga
     2941 gtggcgtgaa tggggaactg cgcttcccaa ataatgccca cctcgagcag atccggctgc
     3001 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     3061 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     3121 cggataattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3181 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     3601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3841 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     3901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4501 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4981 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5161 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     5221 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     5281 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5341 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5401 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     5461 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     5521 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     5581 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     5641 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     5701 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     5761 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     5821 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     5881 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     5941 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     6001 cgggtcctca acgacaggag
//
