LOCUS       sequence-254656-        5443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses  S157A mutant of PfeE in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5443)
  AUTHORS   Perraud Q, Moynie L, Gasser V, Munier M, Godet J, Hoegy F, Mely Y, 
            Mislin GLA, Naismith JH, Schalk IJ
  TITLE     A Key Role for the Periplasmic PfeE Esterase in Iron Acquisition via
            the Siderophore Enterobactin in Pseudomonas aeruginosa.
  JOURNAL   ACS Chem Biol. 2018 Sep 21;13(9):2603-2614. doi: 
            10.1021/acschembio.8b00543. Epub 2018 Aug 22.
  PUBMED    30086222
REFERENCE   2  (bases 1 to 5443)
  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: 
            "10.1021/acschembio.8b00543"; journalName: "ACS Chem Biol"; date: 
            "2018-09-21- 21"; volume: "13"; issue: "9"; pages: "2603-2614"
COMMENT     Sequence Label: pBADHISTEV_PfeES157A
FEATURES             Location/Qualifiers
     source          1..5443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             332..349
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             371..391
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1301..1318)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1471..1557
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1649..1676
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1695..1786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1787..1855,1856..2647)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2005..2024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2689..3144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2776..2795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2986..3007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3255..3843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3744..3763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3997..4014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4180..4202
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4340..4359
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4539..5417)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcttacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcaggag gaatacacca tgagctacta ccatcaccat caccatcacg attacgacat
      361 cccaacgacc gaaaacctgt attttcaggg cgccatgaat ccggaccccg aggcgaccat
      421 ggaccgcagc ctgctgcaac gccaggacct gccctatcgc ttcagcgccg tggacctgga
      481 ctccgtcgat ggccagcggc attaccggct gtggctcggc cgaccgctgc aggcaccgcc
      541 cgccgccggc tacccggtgg tctggatgct cgacggcaac gccgccgtcg gcgccctcga
      601 cgagtcaacc ctgagacgcc tggccgacgg cgacgcaccg ctactggtcg ccatcggcta
      661 ccgcacgccg ctgcgcatcg accgcgccgg gcgtaccttc gactacaccc ccgcgagtcc
      721 cggccaggcc gatcagcgcg acccgctcaa cggcctgccc agcggtggcg ccgacgcctt
      781 cctcgacctg ctgcgcgacg gcatgcgccc ggccgtggcg gcgcaggcgc ctctggacac
      841 tgcgcgacaa accctctggg gacacgccta cggcggcttg ctggtgctcc acgcactgtt
      901 cacccgtccc ggcgagttcg cccgctacgc cgcggccagc ccgtcgctgt ggtggcgcga
      961 cggggcgatc ctcggcgaac gagcggggct cgaacagcgc ctgcgcggaa agcgcgcgga
     1021 actcctgctc tggcgcggca gcgccgaacc ggccagcccg cgcggctcgc tgaaagccga
     1081 gccgggccag gcgatggcgc ggctggtaga tgacctgcgc cgggttgccg ggctcacgct
     1141 ggacttccag ccgctcgacg gcctgggtca tggcgaaacg ctgggggcct cgttgcgcct
     1201 tctgctcgcc cgtccggcgg tggaacgcca gcgctgagga tccgaattcg agctccgtcg
     1261 acaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     1321 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     1381 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     1441 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     1501 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     1561 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     1621 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     1681 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1741 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1801 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1861 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1921 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1981 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     2041 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2101 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2161 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2221 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2281 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2341 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2401 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2461 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2521 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2581 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2641 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
     2701 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2761 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2821 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2881 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     2941 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3001 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     3061 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3121 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
     3181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3361 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3901 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3961 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4021 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4081 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4141 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4201 ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc
     4261 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     4321 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4381 cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
     4441 atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
     4501 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
     4561 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     4621 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     4681 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     4741 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     4801 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     4861 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     4921 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     4981 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     5041 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     5101 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     5161 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     5221 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     5281 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     5341 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     5401 ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
