LOCUS       sequence-434437-        7332 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 5 (OL5) by examining the 
            variant lacking OL5 in its interaction with exogenous peptides..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7332)
  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 7332)
  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-OL5del
FEATURES             Location/Qualifiers
     source          1..7332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1590..1612
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1627..1651
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(1651..1670)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1652..1670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1758..1777)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1983..2060
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2061..3143
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2080..2099)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3715..3906
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3900..3922)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4008..4147
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4063..4082
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4162..4179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4333..4921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4413..4432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5032..5051)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5043..5858
                     /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             5043..5858
                     /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(5116..5135)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5950..6405)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6087..6108)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6299..6318
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(6442..6489)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6485..6503
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(6566..6589)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(6614..6631)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 tgttcaggcc gccgatcacc aggctgacca ggatgatcac cttgccggcc cgcaccacga
       61 agccgcgcag gcgcgaccag gtctgcagca gcaggctctt aaggtgtggc acatggtaca
      121 gcggcagttc catgacgaac ggcgaggcct cgccgcgcat caaggtgtgc ttgagcaaca
      181 gcccggtgag gatcgccacg acgatgccga gcaggtacag ggagaagatc accagcgcgc
      241 cgccctggcc gaagaaggcc ccggcgaata cggcgaagat cgccaggcgc gcgccgcagg
      301 acatgaacgg cgccatcatg atggtaatca gccgctcgcg ctgcgcgtcc agggtccgcg
      361 cgcccatgat cgacggtacg ttgcagccga aaccgacgat cagcggcacg aacgacttgc
      421 ccggcaggcc cagcgcctgc atcaggcggt ccatgacgaa ggccgcgcgt gccatgtagc
      481 cggaatcctc gagcagcgag aggaacaggt acatcaagcc gatctgcggc accagcggca
      541 acaccgtgtt gaccccgcca ccgatgccct gggcgaggaa caccgtgagc caatcgggca
      601 ggccgaagcg gatacccagc cactggatgc cgtcgatgaa gatcgccgac gagcccttgt
      661 cgaagatcgg ctgtagcgcc ccgccgatgt tgatggcgaa gaagaacatc aggtacatca
      721 ccagcaggaa gatcggcaga cccagccagc gattcaggac gacccggtcg agccattggg
      781 tcagccggtg cggctgagcc tgctggtggt cgcagaccgc cgcacagatc tcgccgatca
      841 gccggtagcg cgcatcgacg atcgccagct cgggctcctc gccgcagccc cggcgcgcct
      901 gttccagggt ggcgggcggc aagcccagcg ccgggccgtt gaagatgtcg ccttccagcg
      961 cctgcaacgc cagccagcgc ggctcgatgg ccgaggccgc cggagcgcgc gtttccagaa
     1021 ggtagccgac ctgcgcctgg atggccggtg ggtaatccac cgccagcgcc gcctgcggga
     1081 gctgtaggga gtcgatggcc gccttcagtt cgtcgatgcc atcggcgcgg gtggatacca
     1141 gcggcaccac cgggcagccg agacgccgcg ccagaccgtc gatatcgatg cggatgcgct
     1201 ggctacgggc gatgtcgagc atgttcagcg cgacgatgca ggggatgccc atctcccgca
     1261 gttgcaccgt gaggtacagg ttgcgctcca ggttcgccgc atccaccacg ttgaccagca
     1321 cgtccacttc gccgctggcg atgtagcggc aggcgatctg ctcgtccagc gaggcctggg
     1381 cggaaacgct ggtcaacgaa taggtgccgg gcaggtcgac caggcgcacc gcgtgacgga
     1441 cggtatggaa cgcaccctcc ttgcgctcca cggtgacgcc ggcccagttg ccgacccgct
     1501 ggcgcgagcc ggtgagctgg ttgaacaggg tggtcttgcc ggagttggga ttgccgatca
     1561 ggccgagggt caatgcggtc atatgtatat ctccttctta aagttaaaca aaattatttc
     1621 tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg
     1681 atcgagatcg atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     1741 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     1801 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     1861 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     1921 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc
     1981 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     2041 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     2101 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     2161 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     2221 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     2281 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     2341 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     2401 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     2461 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     2521 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     2581 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     2641 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     2701 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     2761 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     2821 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg
     2881 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     2941 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     3001 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     3061 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3121 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     3181 gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     3241 gcgcggggca tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     3301 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     3361 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     3421 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     3481 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     3541 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     3601 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     3661 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     3721 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     3781 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     3841 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     3901 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     3961 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4021 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4081 tactgagagt gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4141 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4201 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4261 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4741 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4921 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4981 agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     5041 ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg
     5101 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     5161 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     5221 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     5281 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     5341 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     5401 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     5461 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     5521 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     5581 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     5641 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     5701 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     5761 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     5821 aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac
     5881 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5941 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     6001 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     6061 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     6121 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6181 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6241 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6301 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6361 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccaatcc
     6421 ggatatagtt cctcctttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     6481 ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt
     6541 tagcagccta ggtattaatc aattagtggt ggtggtggtg gtggtggtgc tcgagtgcgg
     6601 ccgcaagctt cccactaccg cgtggcacca ggtgacatcc tccctgcccg ctcggggcat
     6661 cggccattcg gcctggcagt tgcagtacct gctcacggcc gaacagacgc aggcaggtca
     6721 tcaggatcag gttgaagagc agcaccacgg cgatggtcag gacgctacgc tcggggtgga
     6781 cctggtagca gagggtcgcc agcgagtagg cgacgttcag cccccagagc accgagaaag
     6841 ccatccagcc cttgctgctt tcccgggcga tcgcgcccat ggcggtcacg cagggcacgt
     6901 agagcaggac gaagaccaga tagctgtagg ccgcgatggg gctgccgaac ttgctcgcca
     6961 tggtgcccat cgagccggtc tccatgtcgc cgtcggccat gctcgcctcc accgggttgg
     7021 ccagcacgct gaggctgaag gtatcgacca ggccgtccca ggtctcgacc agggcctcgc
     7081 gcagttgccc gggcaggtcg tagccttcgt agtcgaaggc ctcgccctgg atctgctcgg
     7141 cggtgtagag ggtgttcagg gtgccgacca ccacttcctt ggccatcgcc ccggtgacca
     7201 ggccgacggt ggcctgccag ttgtccggct gcaccccgag cggcgccagc agcggagtga
     7261 gacgctggct gacgctggcc agcgcggaat ggccgatgtc gccctgcacc ggcttgccgt
     7321 cgagggtgat gc
//
