LOCUS       sequence-434441-        4499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Study the interaction of FeoB in P. aeruginosa, specifically 
            focusing on the role of the outer loop 2 (OL2) by examining the 
            variant lacking OL2 in its interaction with exogenous peptides..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4499)
  AUTHORS   Sharifian Gh M, Norouzi F, Sorci M, Zaidi TS, Pier GB, Achimovich A,
            Ongwae GM, Liang B, Ryan M, Lemke M, Belfort G, Gadjeva M, Gahlmann 
            A, Pires MM, Venter H, Harris TE, Laurie GW
  TITLE     Lacritin cleavage-potentiated targeting of iron - respiratory 
            reciprocity promotes bacterial death.
  JOURNAL   J Biol Chem. 2025 May;301(5):108455. doi: 10.1016/j.jbc.2025.108455.
            Epub 2025 Mar 26.
  PUBMED    40154612
REFERENCE   2  (bases 1 to 4499)
  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: "J Biol 
            Chem."; date: "2025-05"; volume: "301(5):108455. doi"; pages: " 
            10.1016/j.jbc.2025.108455. Epub 2025 Mar 26"
COMMENT     Sequence Label: pET41-FeoB-OL2del
FEATURES             Location/Qualifiers
     source          1..4499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(540..995)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(677..698)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     889..908
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1032..1079)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1075..1093
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1156..1179)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2804..2995
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2989..3011)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3097..3236
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3152..3171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3251..3268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3422..4010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3502..3521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4121..4140)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(4132..4499,1..448)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             join(4132..4499,1..448)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4205..4224)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
       61 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
      121 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
      181 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
      241 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
      301 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
      361 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
      421 tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat
      481 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
      541 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      601 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      661 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      721 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
      781 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      841 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      901 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
      961 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccaatcc ggatatagtt
     1021 cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt
     1081 tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt tagcagccta
     1141 ggtattaatc aattagtggt ggtggtggtg gtggtggtgc tcgagtgcgg ccgcaagctt
     1201 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct atgcgaaaac gcgggaaaaa
     1261 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     1321 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     1381 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     1441 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     1501 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     1561 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     1621 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     1681 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     1741 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     1801 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     1861 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     1921 accgagtccg ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa
     1981 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     2041 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     2101 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     2161 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2221 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc
     2281 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2341 gactagcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     2401 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     2461 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     2521 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     2581 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     2641 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     2701 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     2761 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     2821 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     2881 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     2941 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3001 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3061 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     3121 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     3181 caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3301 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3361 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3601 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3841 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4081 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     4141 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
     4201 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     4261 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     4321 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     4381 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg
     4441 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgc
//
