LOCUS       sequence-342023-        7000 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Complementation plasmid for LetAB without LetB TM region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7000)
  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 7000)
  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: pBEL2039
FEATURES             Location/Qualifiers
     source          1..7000
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        89..107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             139..161
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4046..4064)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4060..4107
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4152..4170)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4238..4342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4343..4411,4412..5203)
                     /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(4561..4580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5374..5962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5863..5882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6116..6133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6148..6287
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6213..6232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6373..6395
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6389..6580)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
       61 gctgcccgag atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg
      121 tttccctcta gaaataattt tgtttaactt taagaaggag atatacatat ggctcttaac
      181 acaccacaaa ttacgccgac aaaaaagata acagtgaggg caatcggcga ggaactgccg
      241 cgtggtgatt accaacgttg cccgcaatgt gacatgctgt ttagcctgcc cgagataaat
      301 tctcatcaaa gtgcctattg tccgcgctgt caggcaaaaa ttcgtgacgg gcgcgactgg
      361 tcgctaacgc gcctggcggc aatggccttc accatgctgt tgcttatgcc gtttgcctgg
      421 ggcgaaccgc tgttgcatat ctggctgtta ggcattcgta ttgacgccaa cgttatgcaa
      481 ggcatctggc aaatgaccaa acagggcgat gcgataacgg ggtcgatggt ctttttctgc
      541 gttattggtg ccccgctcat tctggtgacc tccatagctt atttatggtt tggtaaccga
      601 ctgggaatga atttacgtcc ggtactgctg atgcttgagc gacttaaaga gtgggtaatg
      661 ctcgatatct acctggtcgg cattggcgtt gcttctataa aggtacagga ttatgcccat
      721 atccaggctg gtgtcggctt gttctctttt gtggcgttgg tgattttaac gacggtgacg
      781 ttgtcacatc ttaatgtcga agaactgtgg gagcgatttt atccgcagcg ccccgctacg
      841 cgtagggacg agaaacttcg tgtctgccta gggtgccatt ttaccggcta tccagatcag
      901 cgtggtcgct gcccgcgttg ccatatcccg ctacgcctgc gtcgccgtca tagtctgcaa
      961 aaatgctggg cggcgctgtt agcgtcaatc gttttattgt tacctgccaa cctgttgcct
     1021 atttctatca tttatctgaa cggaggacgg caggaagata caattctttc cggaattatg
     1081 tcgctggcaa gtagcaacat tgcggttgca ggaatcgtgt ttatcgccag tattctggtc
     1141 ccgtttacta aagtgatcgt catgttcact ttactgttga gcattcattt taaatgccag
     1201 caaggtttac gcacacgcat tctgttactg cggatggtga cctggattgg tcgctggtcg
     1261 atgctcgacc tgtttgtcat atctttaacc atgtcgctga ttaatcgcga tcagatcctc
     1321 gcttttacta tgggaccggc tgcgttttat ttcggcgcag cggtaatttt gactattctt
     1381 gctgtggaat ggctggacag ccgcttactt tgggatgcac acgaaagcgg caatgcgcgt
     1441 tttgatgatt gatttcagga gaaccgacgc atgcaggacc ggggtaatac cgtcaccatc
     1501 gactttatgt cggcggatgg tattgttcca ggccgtacgc ctgttcgtta tcagggcgtt
     1561 gaagtcggaa cagtgcagga catcagcctc agcgacgatc ttcgtaagat tgaagtcaag
     1621 gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag agactcagtt ctggctggtg
     1681 acgccaaaag catcgttggc aggtgtctcc gggctggacg ccctcgtcgg tgggaactat
     1741 atcggcatga tgcccgggaa aggtaaagag caggatcact ttgtcgcact cgatacccaa
     1801 ccgaaatatc ggctggacaa tggcgatctg atgatccacc tgcaagcccc cgatctcggt
     1861 tcgctgaaca gcggttcatt ggtctatttc cgcaagatcc cggtgggaaa agtctacgac
     1921 tatgccatca atcccaacaa gcaaggcgtg gtgattgatg tcctgatcga gcggcgtttt
     1981 accgatctgg tgaaaaaagg tagccgtttc tggaacgttt ccggcgttga tgccaacgtc
     2041 agtatcagtg gcgcgaaggt gaaactggaa agcttggcgg cactggttaa cggtgcgatt
     2101 gccttcgatt caccagaaga gtcgaaacct gccgaggcgg aagatacctt tggtctgtat
     2161 gaagatctgg cccacagcca gcgtggcgta ataataaaac tggaactgcc gagtggggcc
     2221 ggattaaccg ccgactcgac gccgttaatg tatcaggggc tggaagtcgg acagctgact
     2281 aaactggatt taaatcctgg tggtaaagtc acaggggaaa tgaccgttga tcccagcgtc
     2341 gttaccctgc ttcgtgaaaa tacccgcatc gaattacgca acccgaaatt atcccttagc
     2401 gatgctaatc tcagcgccct gctgaccggc aaaaccttcg agctggtgcc cggcgatggc
     2461 gagccacgca aagaattcgt tgttgtgcca ggcgaaaaag cactgctgca tgaacctgat
     2521 gttctgacgc tgaccctgac cgcgccggaa agttacggta ttgatgcggg tcagccgctc
     2581 attcttcacg gcgtgcaggt aggccaggtt atcgatcgta aactcaccag caaaggcgtc
     2641 acctttaccg tcgccatcga gcctcagcat cgcgaactgg taaaaggcga tagcaaattt
     2701 gtcgtcaaca gccgtgtcga tgtgaaggtg gggctggatg gcgttgagtt tctcggtgcc
     2761 agcgcctcag aatggatcaa tggcgggatc cgtattctgc cgggcgataa aggtgagatg
     2821 aaagccagct atccactgta tgccaatttg gaaaaagcgc tggagaacag ccttagcgat
     2881 ttacccacca caaccgtgag tttgagtgca gagacgctgc cggatgtgca ggcaggatcg
     2941 gtagtgctgt accgtaaatt tgaagttggt gaagtgatta ccgtgcgtcc gcgagctaac
     3001 gcgtttgaca tcgatctgca tattaagccg gagtatcgca accttctgac cagcaatagc
     3061 gtgttctggg cagaaggcgg ggcgaaagtt cagctgaatg gtagtggtct gaccgtacag
     3121 gcatccccgc tctccagagc tctaaaggga gccattagct tcgacaacct cagcggtgcc
     3181 agcgccagtc agcgtaaagg cgataagcgt attctgtatg cttccgaaac agcggcccgt
     3241 gcggttggcg ggcagattac gcttcacgct ttcgatgccg gaaaactggc ggtcgggatg
     3301 ccaattcgct atctcggtat tgacatcggg caaatccaga cgctggatct gattaccgca
     3361 cgcaatgaag tacaggcaaa ggcggtactt tacccggaat atgtccagac ctttgctcgc
     3421 ggtggtacgc gcttctcagt ggtcacaccg caaatttcgg cagctggcgt tgagcatctt
     3481 gatactatcc tccagccgta tatcaacgtc gaaccaggcc ggggcaatcc tcgccgcgac
     3541 tttgaattac aagaggccac cattactgat tcgcggtacc tggatggctt aagcattatt
     3601 gttgaagcgc cggaagccgg ttcgttaggc atcggtacgc ctgtgctgtt ccgtggtctg
     3661 gaagtcggta cggttacagg aatgacgctg gggacattgt cagatcgcgt gatgattgcg
     3721 atgcgcatca gtaaacgcta tcaacacctg gtgcgtaaca attccgtctt ctggttggca
     3781 tcgggttaca gtctggactt tggtctgacg ggcggcgtag tgaaaaccgg cacctttaac
     3841 cagtttatcc gtggcggcat cgccttcgcc acgcctccgg gtacgccact ggcaccgaaa
     3901 gcccaggaag gcaaacactt cctgttgcag gaaagtgaac cgaaagagtg gcgtgaatgg
     3961 ggaactgcgc ttcccaaata atgcccacct cgagcagatc cggctgctaa caaagcccga
     4021 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     4081 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ataattcttg
     4141 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4201 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4261 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4321 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4381 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4441 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4501 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4561 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     4621 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4681 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4741 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4801 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4861 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     4921 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4981 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5041 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5101 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5161 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5221 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5281 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5521 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6001 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6061 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6121 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     6181 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6241 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     6301 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6361 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     6421 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     6481 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     6541 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     6601 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     6661 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     6721 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     6781 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     6841 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     6901 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     6961 tcggtgattc attctgctaa ccagtaaggc aaccccgcca
//
