LOCUS       sequence-310632-        7063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB with LetB TM region replaced by 
            PelB leader sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7063)
  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 7063)
  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"; 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: pBEL2040
FEATURES             Location/Qualifiers
     source          1..7063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     584..603
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        584..602
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             634..656
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     1966..2031
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(4604..4622)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4618..4665
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4710..4728)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4796..4900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4901..4969,4970..5761)
                     /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(5119..5138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5932..6520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6421..6440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6674..6691
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6706..6845
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6771..6790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6931..6953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(6947..7063,1..75))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
       61 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
      121 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
      181 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
      241 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
      301 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
      361 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
      421 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
      481 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
      541 tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata
      601 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
      661 catatggctc ttaacacacc acaaattacg ccgacaaaaa agataacagt gagggcaatc
      721 ggcgaggaac tgccgcgtgg tgattaccaa cgttgcccgc aatgtgacat gctgtttagc
      781 ctgcccgaga taaattctca tcaaagtgcc tattgtccgc gctgtcaggc aaaaattcgt
      841 gacgggcgcg actggtcgct aacgcgcctg gcggcaatgg ccttcaccat gctgttgctt
      901 atgccgtttg cctggggcga accgctgttg catatctggc tgttaggcat tcgtattgac
      961 gccaacgtta tgcaaggcat ctggcaaatg accaaacagg gcgatgcgat aacggggtcg
     1021 atggtctttt tctgcgttat tggtgccccg ctcattctgg tgacctccat agcttattta
     1081 tggtttggta accgactggg aatgaattta cgtccggtac tgctgatgct tgagcgactt
     1141 aaagagtggg taatgctcga tatctacctg gtcggcattg gcgttgcttc tataaaggta
     1201 caggattatg cccatatcca ggctggtgtc ggcttgttct cttttgtggc gttggtgatt
     1261 ttaacgacgg tgacgttgtc acatcttaat gtcgaagaac tgtgggagcg attttatccg
     1321 cagcgccccg ctacgcgtag ggacgagaaa cttcgtgtct gcctagggtg ccattttacc
     1381 ggctatccag atcagcgtgg tcgctgcccg cgttgccata tcccgctacg cctgcgtcgc
     1441 cgtcatagtc tgcaaaaatg ctgggcggcg ctgttagcgt caatcgtttt attgttacct
     1501 gccaacctgt tgcctatttc tatcatttat ctgaacggag gacggcagga agatacaatt
     1561 ctttccggaa ttatgtcgct ggcaagtagc aacattgcgg ttgcaggaat cgtgtttatc
     1621 gccagtattc tggtcccgtt tactaaagtg atcgtcatgt tcactttact gttgagcatt
     1681 cattttaaat gccagcaagg tttacgcaca cgcattctgt tactgcggat ggtgacctgg
     1741 attggtcgct ggtcgatgct cgacctgttt gtcatatctt taaccatgtc gctgattaat
     1801 cgcgatcaga tcctcgcttt tactatggga ccggctgcgt tttatttcgg cgcagcggta
     1861 attttgacta ttcttgctgt ggaatggctg gacagccgct tactttggga tgcacacgaa
     1921 agcggcaatg cgcgttttga tgattgattt caggagaacc gacgcatgaa atacctgctg
     1981 ccgaccgctg ctgctggtct gctgctcctc gctgcccagc cggcgatggc ccaggaccgg
     2041 ggtaataccg tcaccatcga ctttatgtcg gcggatggta ttgttccagg ccgtacgcct
     2101 gttcgttatc agggcgttga agtcggaaca gtgcaggaca tcagcctcag cgacgatctt
     2161 cgtaagattg aagtcaaggt cagcatcaag tccgatatga aagatgcgct gcgcgaagag
     2221 actcagttct ggctggtgac gccaaaagca tcgttggcag gtgtctccgg gctggacgcc
     2281 ctcgtcggtg ggaactatat cggcatgatg cccgggaaag gtaaagagca ggatcacttt
     2341 gtcgcactcg atacccaacc gaaatatcgg ctggacaatg gcgatctgat gatccacctg
     2401 caagcccccg atctcggttc gctgaacagc ggttcattgg tctatttccg caagatcccg
     2461 gtgggaaaag tctacgacta tgccatcaat cccaacaagc aaggcgtggt gattgatgtc
     2521 ctgatcgagc ggcgttttac cgatctggtg aaaaaaggta gccgtttctg gaacgtttcc
     2581 ggcgttgatg ccaacgtcag tatcagtggc gcgaaggtga aactggaaag cttggcggca
     2641 ctggttaacg gtgcgattgc cttcgattca ccagaagagt cgaaacctgc cgaggcggaa
     2701 gatacctttg gtctgtatga agatctggcc cacagccagc gtggcgtaat aataaaactg
     2761 gaactgccga gtggggccgg attaaccgcc gactcgacgc cgttaatgta tcaggggctg
     2821 gaagtcggac agctgactaa actggattta aatcctggtg gtaaagtcac aggggaaatg
     2881 accgttgatc ccagcgtcgt taccctgctt cgtgaaaata cccgcatcga attacgcaac
     2941 ccgaaattat cccttagcga tgctaatctc agcgccctgc tgaccggcaa aaccttcgag
     3001 ctggtgcccg gcgatggcga gccacgcaaa gaattcgttg ttgtgccagg cgaaaaagca
     3061 ctgctgcatg aacctgatgt tctgacgctg accctgaccg cgccggaaag ttacggtatt
     3121 gatgcgggtc agccgctcat tcttcacggc gtgcaggtag gccaggttat cgatcgtaaa
     3181 ctcaccagca aaggcgtcac ctttaccgtc gccatcgagc ctcagcatcg cgaactggta
     3241 aaaggcgata gcaaatttgt cgtcaacagc cgtgtcgatg tgaaggtggg gctggatggc
     3301 gttgagtttc tcggtgccag cgcctcagaa tggatcaatg gcgggatccg tattctgccg
     3361 ggcgataaag gtgagatgaa agccagctat ccactgtatg ccaatttgga aaaagcgctg
     3421 gagaacagcc ttagcgattt acccaccaca accgtgagtt tgagtgcaga gacgctgccg
     3481 gatgtgcagg caggatcggt agtgctgtac cgtaaatttg aagttggtga agtgattacc
     3541 gtgcgtccgc gagctaacgc gtttgacatc gatctgcata ttaagccgga gtatcgcaac
     3601 cttctgacca gcaatagcgt gttctgggca gaaggcgggg cgaaagttca gctgaatggt
     3661 agtggtctga ccgtacaggc atccccgctc tccagagctc taaagggagc cattagcttc
     3721 gacaacctca gcggtgccag cgccagtcag cgtaaaggcg ataagcgtat tctgtatgct
     3781 tccgaaacag cggcccgtgc ggttggcggg cagattacgc ttcacgcttt cgatgccgga
     3841 aaactggcgg tcgggatgcc aattcgctat ctcggtattg acatcgggca aatccagacg
     3901 ctggatctga ttaccgcacg caatgaagta caggcaaagg cggtacttta cccggaatat
     3961 gtccagacct ttgctcgcgg tggtacgcgc ttctcagtgg tcacaccgca aatttcggca
     4021 gctggcgttg agcatcttga tactatcctc cagccgtata tcaacgtcga accaggccgg
     4081 ggcaatcctc gccgcgactt tgaattacaa gaggccacca ttactgattc gcggtacctg
     4141 gatggcttaa gcattattgt tgaagcgccg gaagccggtt cgttaggcat cggtacgcct
     4201 gtgctgttcc gtggtctgga agtcggtacg gttacaggaa tgacgctggg gacattgtca
     4261 gatcgcgtga tgattgcgat gcgcatcagt aaacgctatc aacacctggt gcgtaacaat
     4321 tccgtcttct ggttggcatc gggttacagt ctggactttg gtctgacggg cggcgtagtg
     4381 aaaaccggca cctttaacca gtttatccgt ggcggcatcg ccttcgccac gcctccgggt
     4441 acgccactgg caccgaaagc ccaggaaggc aaacacttcc tgttgcagga aagtgaaccg
     4501 aaagagtggc gtgaatgggg aactgcgctt cccaaataat gcccacctcg agcagatccg
     4561 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     4621 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     4681 atatccggat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4741 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4801 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4861 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4921 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4981 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5041 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5101 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5161 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5221 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5281 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5341 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5401 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5461 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5521 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5581 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5641 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5701 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5761 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5821 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5881 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5941 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6001 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6061 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6121 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6181 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6241 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6301 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6361 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6421 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6481 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6541 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6601 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6661 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     6721 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     6781 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     6841 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6901 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6961 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     7021 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gag
//
