LOCUS       sequence-434055-        7320 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 1 (OL1) by examining the 
            variant lacking OL1 in its interaction with exogenous peptides..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7320)
  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 7320)
  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-OL1del
FEATURES             Location/Qualifiers
     source          1..7320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1900..1917
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1942..1965
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2028..2046)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2042..2089
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2126..2581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2213..2232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2423..2444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2673..3488)
                     /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             complement(2673..3488)
                     /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     3396..3415
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3480..3499
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3610..4198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4099..4118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4352..4369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4384..4523
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4449..4468)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4609..4631
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4625..4816)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5388..6470)
                     /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     6432..6451
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6471..6548)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6754..6773
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6861..6880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6861..6879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6880..6904
                     /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)."
     RBS             6919..6941
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 gcccgtagcc agcgcatccg catcgatatc gacggtctgg cgcggcgtct cggctgcccg
       61 gtggtgccgc tggtatccac ccgcgccgat ggcatcgacg aactgaaggc ggccatcgac
      121 tccctacagc tcccgcaggc ggcgctggcg gtggattacc caccggccat ccaggcgcag
      181 gtcggctacc ttctggaaac gcgcgctccg gcggcctcgg ccatcgagcc gcgctggctg
      241 gcgttgcagg cgctggaagg cgacatcttc aacggcccgg cgctgggctt gccgcccgcc
      301 accctggaac aggcgcgccg gggctgcggc gaggagcccg agctggcgat cgtcgatgcg
      361 cgctaccggc tgatcggcga gatctgtgcg gcggtctgcg accaccagca ggctcagccg
      421 caccggctga cccaatggct cgaccgggtc gtcctgaatc gctggctggg tctgccgatc
      481 ttcctgctgg tgatgtacct gatgttcttc ttcgccatct tcgacaaggg ctcgtcggcg
      541 atcttcatcg acggcatcca gtggctgggt atccgcttcg gcctgcccga ttggctcacg
      601 gtgttcctcg cccagggcat cggtggcggg gtcaacacgg tgttgccgct ggtgccgcag
      661 atcggcttga tgtacctgtt cctctcgctg ctcgaggatt ccggctacat ggcacgcgcg
      721 gccttcgtca tggaccgcct gatgcaggcg ctgggcctgc cgggcaagtc gttcgtgccg
      781 ctgatcgtcg gtttcggctg caacgtaccg tcgatcatgg gcgcgcggac cctggacgcg
      841 cagcgcgagc ggctgattac catcatgatg gcgccgttca tgtcctgcgg cgcgcgcctg
      901 gcgatcttcg ccgtattcgc cggggccttc ttcggccagg gcggcgcgct ggtgatcttc
      961 tccctgtacc tgctcggcat cgtcgtggcg atcctcaccg ggctgttgct caagcacacc
     1021 ttgatgcgcg gcgaggcctc gccgttcgtc atggaactgc cgctgtacca tgtgccacac
     1081 cttaagagcc tgctgctgca gacctggtcg cgcctgcgcg gcttcgtggt gcgggccggc
     1141 aaggtgatca tcctggtcag cctggtgatc ggcggcctga acagcatcac cctcgacggc
     1201 aagccggtgc agggcgacat cggccattcc gcgctggcca gcgtcagcca gcgtctcact
     1261 ccgctgctgg cgccgctcgg ggtgcagccg gacaactggc aggccaccgt cggcctggtc
     1321 accggggcga tggccaagga agtggtggtc ggcaccctga acaccctcta caccgccgag
     1381 cagatccagg gcgaggcctt cgactacgaa ggctacgacc tgcccgggca actgcgcgag
     1441 gccctggtcg agacctggga cggcctggtc gataccttca gcctcagcgt gctggccaac
     1501 ccggtggagg cgagcatggc cgacggcgac atggagaccg gctcgatggg caccatggcg
     1561 agcaagttcg gcagccccat cgcggcctac agctatctgg tcttcgtcct gctctacgtg
     1621 ccctgcgtga ccgccatggg cgcgatcgcc cgggaaagca gcaagggctg gatggctttc
     1681 tcggtgctct gggggctgaa cgtcgcctac tcgctggcga ccctctgcta ccaggtcgcc
     1741 accttcgccg cgcaccccga gcgtagcgtc ctgaccatcg ccgtggtgct gctcttcaac
     1801 ctgatcctga tgacctgcct gcgtctgttc ggccgtgagc aggtactgca actgccaggc
     1861 cgaatggccg atgccccgag cgggcaggga ggatgtcacc tggtgccacg cggtagtggg
     1921 aagcttgcgg ccgcactcga gcaccaccac caccaccacc accactaatt gattaatacc
     1981 taggctgcta aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     2041 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     2101 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2161 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2221 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2281 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2341 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2401 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2461 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2521 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2581 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2641 attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga
     2701 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2761 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2821 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2881 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     2941 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     3001 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     3061 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     3121 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc
     3181 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     3241 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     3301 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3361 aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3421 aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata
     3481 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac
     3541 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3601 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3661 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3721 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3781 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3841 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3901 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3961 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4021 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4081 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4141 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4201 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4261 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4321 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4381 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     4441 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     4501 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     4561 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4621 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     4681 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     4741 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     4801 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     4861 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     4921 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     4981 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     5041 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     5101 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     5161 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     5221 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgctagtca
     5281 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     5341 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     5401 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5461 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     5521 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     5581 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     5641 tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta
     5701 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     5761 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     5821 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     5881 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     5941 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     6001 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     6061 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     6121 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     6181 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     6241 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     6301 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     6361 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     6421 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     6481 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     6541 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     6601 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     6661 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     6721 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     6781 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     6841 cgatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     6901 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat gaccgcattg
     6961 accctcggcc tgatcggcaa tcccaactcc ggcaagacca ccctgttcaa ccagctcacc
     7021 ggctcgcgcc agcgggtcgg caactgggcc ggcgtcaccg tggagcgcaa ggagggtgcg
     7081 ttccataccg tccgtcacgc ggtgcgcctg gtcgacctgc ccggcaccta ttcgttgacc
     7141 agcgtttccg cccaggcctc gctggacgag cagatcgcct gccgctacat cgccagcggc
     7201 gaagtggacg tgctggtcaa cgtggtggat gcggcgaacc tggagcgcaa cctgtacctc
     7261 acggtgcaac tgcgggagat gggcatcccc tgcatcgtcg cgctgaacat gctcgacatc
//
