LOCUS       sequence-254679-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Pseudomonas aeruginosa PirA in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  AUTHORS   Moynie L, Luscher A, Rolo D, Pletzer D, Tortajada A, Weingart H, 
            Braun Y, Page MG, Naismith JH, Kohler T
  TITLE     Structure and Function of the PiuA and PirA Siderophore-Drug 
            Receptors from Pseudomonas aeruginosa and Acinetobacter baumannii.
  JOURNAL   Antimicrob Agents Chemother. 2017 Mar 24;61(4). pii: AAC.02531-16. 
            doi: 10.1128/AAC.02531-16. Print 2017 Apr.
  PUBMED    28137795
REFERENCE   2  (bases 1 to 5845)
  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"; journalName: "Antimicrob 
            Agents Chemother."; date: "2017-03-24"; volume: "61(4). pii: 
            AAC.02531-16. doi"; pages: " 10.1128/AAC.02531-16. Print 2017 Apr"
COMMENT     Sequence Label: pET20b_Pa-PirA
FEATURES             Location/Qualifiers
     source          1..5845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2158..2175
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2200..2217
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2270..2288)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2284..2331
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2368..2823
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2455..2474)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2665..2686
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2849..2953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2954..3022,3023..3814)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3172..3191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3985..4573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4474..4493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4727..4744
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4759..4898
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4824..4843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4984..5006
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5000..5191)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5700..5719
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5700..5718
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5750..5772
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     5780..5845
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
ORIGIN
        1 atggatatcg aggatgagcg tctggaagaa ctggacgagc gtgcggagag cgttgtgcag
       61 ctgggtgacg aggtcgtttt gggtaccgcg gaacaagagt tgaaacaggc cccaggtgtt
      121 agcatcatta ccgccgaaga tattcgcaaa cgtccgccgg tcaacgatct gagcgagatt
      181 atccgtacca tgcctggtgt gaatttgacc ggcaatagca gcagcggcca acgtggtaat
      241 aaccgccaaa ttgacattcg cggcatgggt ccggaaaaca ctctgatcct ggttgatggc
      301 aaaccggtca gcagccgtaa tagcgtacgt tacggctggc gtggcgaacg tgacacccgc
      361 ggtgacagca actgggttcc gccggaagaa gtcgagcgta tcgaggttct gcgtggtccg
      421 gcagctgctc gctatggtag cggcgctgcg ggtggtgtgg tcaatatcat cacgaagcgc
      481 ccgaccgatc gtttgcgcgg tagcatgacg gtgttcacca acatcccgga gagctcgaag
      541 gacggtgcga cccgccgtgc gaactttagc ctgagcggtc cgctgaccga agcactgagc
      601 tttcgtgcgt acggcagcgc aaataagacc gactctgacg atacggacat caacttgggt
      661 cacacggtga acccgagccg caccgtggct ggtcgtgaag gtgtccgtaa tcgtgatctg
      721 tccggcatgc tgtcttggca agtcaccccg gatcaggttg tggattttga ggccggcttc
      781 agccgtcaag gtaatatcta cgcgggtgac acgcagaata acaatggcac cgccaatacc
      841 cagggcctgg cagacgatgg tgcggagact aatcgtatgt atcgcgaaaa ctatgcgatt
      901 acccacaacg gtacctggag cttcggcacc tcgcgcttcg ttgcgcagta cgactccacc
      961 cgcaataatc gtttggaaga gggtctggcg ggttctgtcg aaggtcagat tggtgcagat
     1021 cgcagcttta gcgcaagcaa gctggaaaac tatcgtctgt cgggtgagct gaatttgcca
     1081 ctgcacgcgc tgtttgagca agtgctgact gtgggtgccg agtggaataa agaaaccctg
     1141 aacgacccat ctagcctgaa acagggcttc gtgggtagcg acagcctgcc gggtacgcct
     1201 gcggcaggca gccgcagccc gaagtcgaag gcggagatcc gtgcgctgta cgttgaggat
     1261 aacattgaac tgcgtccggg caccatgctg actccgggtc tgcgtctgga tgatcatagc
     1321 gacttcggtc tgaactggag cccaagcctg aatgcgtccc agacgctggg tgagtatttc
     1381 accgtaaagg caggcattgc tcgtgcattc aaagcaccga acctgtacca aagcaaccct
     1441 aactatttgc tgtacacgcg tggcaacggc tgcccgattc agacgagctc tggcggttgt
     1501 tatctggtgg gtaatgaaaa cctggacgcg gaaacgagcg tcaacaaaga gctgggtatc
     1561 gagttccgcc gtgacggttg ggtcgcgggt ctgacctact ttcgtaacga ctataagaac
     1621 aagatcgttg caccgctgga tgttatgggc cagaccggca cgggcaataa cattctgcag
     1681 tggagcaacg cgaagaaggc ggtcgtcgaa ggcctggaag gcaacttgct ggttccgttg
     1741 catgaggacc tgtcttggag caccaacctg acgtacatgc tgcagtcgaa agacaaagat
     1801 acgggtaatc cgctgtccgt tattccggag tacacgctga atagcaccct ggactggcaa
     1861 gcctcggagc gtctgtccac gcaactgacc agcacgatct acggtcgtca agaaccgccg
     1921 aaacacggta ccagccgcaa cactccggtt gtgtcccgca aagaggtggg tacctacggc
     1981 atttggggcg tttccgctgg ctacacgttt agcgagaatc tgagcgtgcg tggcggtgtg
     2041 agcaatctgt ttgataaacg tctgtatcgc caaggtaaca gcttcgacgc cggtgccgcc
     2101 acctacaatg agccgggtcg tgcatactat gtttccatga cgaccagctt tggatcccat
     2161 caccaccatc atcactaaaa gcttgcggcc gcactcgagc accaccacca ccaccactga
     2221 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2281 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     2341 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     2401 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2461 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2521 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2581 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2641 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2701 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2761 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     2821 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2881 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2941 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3001 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3061 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3121 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3181 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3241 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3301 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3361 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3421 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3481 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3541 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3601 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3661 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3721 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3781 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3841 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3901 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3961 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4021 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4081 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     4141 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4201 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4261 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4321 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4381 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4441 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4501 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4561 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4621 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4681 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4741 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4801 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     4861 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     4921 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4981 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     5041 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     5101 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     5161 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     5221 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     5281 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     5341 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     5401 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     5461 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     5521 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     5581 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     5641 cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatctcgatc ccgcgaaatt
     5701 aatacgactc actataggga gaccacaacg gtttccctct agaaataatt ttgtttaact
     5761 ttaagaagga gatatacata tgaaatacct gctgccgacc gctgctgctg gtctgctgct
     5821 cctcgctgcc cagccggcga tggcc
//
