LOCUS       sequence-447733-        7930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PaFtsH2H1-link-32-6xHis: PaftsH2(ftsH2_F124-V154del_ins 
            ftsH1_M120-T151)-6xHis cloned in XbaI/NheI site in pET-28a(+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7930)
  AUTHORS   Mawla GD, Kamal SM, Cao LY, Purhonen P, Hebert H, Sauer RT, Baker 
            TA, Romling U
  TITLE     The membrane-cytoplasmic linker defines activity of FtsH proteases 
            in Pseudomonas aeruginosa clone C.
  JOURNAL   J Biol Chem. 2024 Feb;300(2):105622. doi: 10.1016/j.jbc.2023.105622.
            Epub 2024 Jan 3.
  PUBMED    38176647
REFERENCE   2  (bases 1 to 7930)
  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: "2024-02"; volume: "300(2):105622. doi"; pages: " 
            10.1016/j.jbc.2023.105622. Epub 2024 Jan 3"
COMMENT     Sequence Label: pSG086
FEATURES             Location/Qualifiers
     source          1..7930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     447..469
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     462..478
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     488..507
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        488..506
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2467..2751)
                     /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     complement(2525..2544)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2778..3656
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(3748..3764)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3749..3765)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3794..3814)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3794..3812)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3833..3849)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3833..3849)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3846..3868)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3857..3873
                     /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)."
     promoter        complement(join(3881..3887,3888..3905,3906..3911))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3926..3947
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4217..4222,4223..4239,4240..4245)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4434..4967
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     rep_origin      6249..7020
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             7021..7683
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
       61 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
      121 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agcaaccagg
      181 cgtttaaggg caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca
      241 gtacggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      301 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
      361 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
      421 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
      481 gcgcgcgtaa tacgactcac tatagggcga attggagctc caccgcggtg gcggccgctc
      541 tagatcatgg tgtggaccct tggggtgtag agagtgaggt ttcgggctgg aggggctccg
      601 acgcgcggga ttcggactgc aagccctgcg tcagttggcg gatgtcgttc tcgtcgagcg
      661 tttcccgcgc cagcagttcg cgcgcacaac gctccagcac cgcgcggttg gtgtcgagga
      721 tccggtaggc gcgctcgaac acgcccatca cgatgtcgcg gatggcctga tcgatgcgcg
      781 cctgggtcga ttcggccacc cggcagccgc cgtgggccag ttccggtgta tcgagaaagc
      841 ggggccgctg cgcctcgaag gcgatgtagc ccagtccttc gtccatgcca aagcgggtga
      901 tcatgtcgcg ggcaatgtcg gtggcccgtg cgaggtcatc ggcagccccg gtggacagct
      961 cgccgaacac gagcttctcg gccgcacgac cgcccagcag cacggcgatc ttgtgttcca
     1021 gatcggcgcg cgtcatcagg aagcggtctt cggtggggcg ttgcagcgtg tagcccagcg
     1081 cgcccatgcc gcgcgggatg atcgagatct tgtgcacggg gtcggtgccg ggcagcgcca
     1141 gcgccaccag cgcatggccc atctcgtgat aggccacggt ttcccgctcc ttgggattga
     1201 gcactcggtt cttcttctcc aggcccgcca cgatgcgctc gatggcggcg gtgaaatcct
     1261 gcaactccac ggcggacgct ttgcgccggg tcgcggccag cgcggcctcg ttgaccaggt
     1321 tcgcgaggtc tgcacccgaa aagcctgtgg tcagcgcagc cacctgttcg agatcgatat
     1381 cctgagccag cgtcaccttc ttgacgtgga ctttcaggat gtccagccgt cccttcttgt
     1441 cgggccggtc caccagcacc tggcggtcga agcggccggc acgcagcagc gcctggtcga
     1501 ggatttcggg gcggttggtg gcggcgagga tgatcagccc caccgagctg tcgaagccgt
     1561 ccatctcggt gagcagctgg ttgagcgtct gctcacgctc gtcgtggccg ccgatgggcc
     1621 cgccgacgcc gcgtgcacgg cccagcgcgt cgagttcatc gatgaagatg atggccggcg
     1681 cctgcccgcg cgcctgctcg aacaggtcgc gcacccgcgc cgcgcccacg ccgacaaaca
     1741 tctcgacgaa ctccgagccg gagatagaga agaacggtac cccggcctcg cccgccacgg
     1801 ccttggccag cagggtcttg cccgtgccgg gcgggccaac cagcaacaca cctttcggga
     1861 tgcgcgcgcc gaggcggccg taatcctgcg ggttcttcag gaagtcgacg atttcgacca
     1921 gctcggcctt ggcctcatcg acacccgcga catcggcaaa ggtggtcttg acctggtcct
     1981 cggacagcag gcgcgccttg ctcttgccga agctcatcgg gccgccgcga ccgccgccgc
     2041 caccctgcat ctggcgcatc aggaagaacc agacgccgaa gaaggccacc gccggcagaa
     2101 tccaggagag cacatcacgt agccaggtgc tttccaccac ccgcgcgtaa ggcacgtcgt
     2161 acttggacag ccgctcggcc aggtcgggtt cgacgcgagt ggccacgatg gtggtcttgc
     2221 cccggctgtc cggcgatttc aggcgcccgg tgaccgtgcg gtccgacacc agcacttcgg
     2281 cgacgcgccc ctcggccagc gccttctcga attcgctgta gggcacgggc tcgacggtct
     2341 tggccgcctg ccagtagttc tgcagcgtca gcagcaacag gccggcgacg atccagtagc
     2401 caatgttcca ttgatctttc ttttccatgg gcaatgttcc tcgcaagcta gcccaaaaaa
     2461 acgggtatgg agaaacagta gagagttgcg ataaaaagcg tcaggtagga tccgctaatc
     2521 ttatggataa aaatgctatg gcatagcaaa gtgtgacgcc gtgcaaataa tcaatgtgga
     2581 cttttctgcc gtgattatag acacttttgt tacgcgtttt tgtcatggct ttggtcccgc
     2641 tttgttacag aatgctttta ataagcgggg ttaccggttt ggttagcgag aagagccagt
     2701 aaaagacgca gtgacggcaa tgtctgatgc aatatggaca attggtttct tctctgaatg
     2761 gcgggagtat gaaaagtatg gctgaagcgc aaaatgatcc cctgctgccg ggatactcgt
     2821 ttaatgccca tctggtggcg ggtttaacgc cgattgaggc caacggttat ctcgattttt
     2881 ttatcgaccg accgctggga atgaaaggtt atattctcaa tctcaccatt cgcggtcagg
     2941 gggtggtgaa aaatcaggga cgagaatttg tttgccgacc gggtgatatt ttgctgttcc
     3001 cgccaggaga gattcatcac tacggtcgtc atccggaggc tcgcgaatgg tatcaccagt
     3061 gggtttactt tcgtccgcgc gcctactggc atgaatggct taactggccg tcaatatttg
     3121 ccaatacggg gttctttcgc ccggatgaag cgcaccagcc gcatttcagc gacctgtttg
     3181 ggcaaatcat taacgccggg caaggggaag ggcgctattc ggagctgctg gcgataaatc
     3241 tgcttgagca attgttactg cggcgcatgg aagcgattaa cgagtcgctc catccaccga
     3301 tggataatcg ggtacgcgag gcttgtcagt acatcagcga tcacctggca gacagcaatt
     3361 ttgatatcgc cagcgtcgca cagcatgttt gcttgtcgcc gtcgcgtctg tcacatcttt
     3421 tccgccagca gttagggatt agcgtcttaa gctggcgcga ggaccaacgt atcagccagg
     3481 cgaagctgct tttgagcacc acccggatgc ctatcgccac cgtcggtcgc aatgttggtt
     3541 ttgacgatca actctatttc tcgcgggtat ttaaaaaatg caccggggcc agcccgagcg
     3601 agttccgtgc cggttgtgaa gaaaaagtga atgatgtagc cgtcaagttg tcataattgg
     3661 taacgaatca gacaattgac ggcttgacgg agtagcatag ggtttgcaga atccctgctt
     3721 cgtccatttg acaggcacat tatgcatcga taccgtcgac ctcgaggggg ggcccggtac
     3781 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3841 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3901 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3961 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4021 gcgcggggag aggcggtttg cgtattgggc gcatgcataa aaactgttgt aattcattaa
     4081 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     4141 tcagcacctt gtcgccttgc gtataatatt tgcccatgga cgcacaccgt ggaaacggat
     4201 gaaggcacga acccagttga cataagcctg ttcggttcgt aaactgtaat gcaagtagcg
     4261 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     4321 ggcggttttc atggcttgtt atgactgttt ttttgtacag tctatgcctc gggcatccaa
     4381 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     4441 gcagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta ggtggctcaa
     4501 gtatgggcat cattcgcaca tgtaggctcg gccctgacca agtcaaatcc atgcgggctg
     4561 ctcttgatct tttcggtcgt gagttcggag acgtagccac ctactcccaa catcagccgg
     4621 actccgatta cctcgggaac ttgctccgta gtaagacatt catcgcgctt gctgccttcg
     4681 accaagaagc ggttgttggc gctctcgcgg cttacgttct gcccaagttt gagcagccgc
     4741 gtagtgagat ctatatctat gatctcgcag tctccggaga gcaccggagg cagggcattg
     4801 ccaccgcgct catcaatctc ctcaagcatg aggccaacgc gcttggtgct tatgtgatct
     4861 acgtgcaagc agattacggt gacgatcccg cagtggctct ctatacaaag ttgggcatac
     4921 gggaagaagt gatgcacttt gatatcgacc caagtaccgc cacctaacaa ttcgttcaag
     4981 ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt gtcacaacgc cgcaggtggc
     5041 acttttcggg gaaatgtgcg cgcccgcgtt cctgctggcg ctgggcctgt ttctggcgct
     5101 ggacttcccg ctgttccgtc agcagctttt cgcccacggc cttgatgatc gcggcggcct
     5161 tggcctgcat atcccgattc aacggcccca gggcgtccag aacgggcttc aggcgctccc
     5221 gaaggtctcg ggccgtctct tgggcttgat cggccttctt gcgcatctca cgcgctcctg
     5281 cggcggcctg tagggcaggc tcatacccct gccgaaccgc ttttgtcagc cggtcggcca
     5341 cggcttccgg cgtctcaacg cgctttgaga ttcccagctt ttcggccaat ccctgcggtg
     5401 cataggcgcg tggctcgacc gcttgcgggc tgatggtgac gtggcccact ggtggccgct
     5461 ccagggcctc gtagaacgcc tgaatgcgcg tgtgacgtgc cttgctgccc tcgatgcccc
     5521 gttgcagccc tagatcggcc acagcggccg caaacgtggt ctggtcgcgg gtcatctgcg
     5581 ctttgttgcc gatgaactcc ttggccgaca gcctgccgtc ctgcgtcagc ggcaccacga
     5641 acgcggtcat gtgcgggctg gtttcgtcac ggtggatgct ggccgtcacg atgcgatccg
     5701 ccccgtactt gtccgccagc cacttgtgcg ccttctcgaa gaacgccgcc tgctgttctt
     5761 ggctggccga cttccaccat tccgggctgg ccgtcatgac gtactcgacc gccaacacag
     5821 cgtccttgcg ccgcttctct ggcagcaact cgcgcagtcg gcccatcgct tcatcggtgc
     5881 tgctggccgc ccagtgctcg ttctctggcg tcctgctggc gtcagcgttg ggcgtctcgc
     5941 gctcgcggta ggcgtgcttg agactggccg ccacgttgcc cattttcgcc agcttcttgc
     6001 atcgcatgat cgcgtatgcc gccatgcctg cccctccctt ttggtgtcca accggctcga
     6061 cgggggcagc gcaaggcggt gcctccggcg ggccactcaa tgcttgagta tactcactag
     6121 actttgcttc gcaaagtcgt gaccgcctac ggcggctgcg gcgccctacg ggcttgctct
     6181 ccgggcttcg ccctgcgcgg tcgctgcgct cccttgccag cccgtggata tgtggacgat
     6241 ggccgcgagc ggccaccggc tggctcgctt cgctcggccc gtggacaacc ctgctggaca
     6301 agctgatgga caggctgcgc ctgcccacga gcttgaccac agggattgcc caccggctac
     6361 ccagccttcg accacatacc caccggctcc aactgcgcgg cctgcggcct tgccccatca
     6421 atttttttaa ttttctctgg ggaaaagcct ccggcctgcg gcctgcgcgc ttcgcttgcc
     6481 ggttggacac caagtggaag gcgggtcaag gctcgcgcag cgaccgcgca gcggcttggc
     6541 cttgacgcgc ctggaacgac ccaagcctat gcgagtgggg gcagtcgaag ggcgaagccc
     6601 gcccgcctgc cccccgagcc tcacggcggc gagtgcgggg gttccaaggg ggcagcgcca
     6661 ccttgggcaa ggccgaaggc cgcgcagtcg atcaacaagc cccggagggg ccactttttg
     6721 ccggaggggg agccgcgccg aaggcgtggg ggaaccccgc aggggtgccc ttctttgggc
     6781 accaaagaac tagatatagg gcgaaatgcg aaagacttaa aaatcaacaa cttaaaaaag
     6841 gggggtacgc aacagctcat tgcggcaccc cccgcaatag ctcattgcgt aggttaaaga
     6901 aaatctgtaa ttgactgcca cttttacgca acgcataatt gttgtcgcgc tgccgaaaag
     6961 ttgcagctga ttgcgcatgg tgccgcaacc gtgcggcacc cctaccgcat ggagataagc
     7021 atggccacgc agtccagaga aatcggcatt caagccaaga acaagcccgg tcactgggtg
     7081 caaacggaac gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa acccacggcg
     7141 gcaatgctgc tgcatcacct cgtggcgcag atgggccacc agaacgccgt ggtggtcagc
     7201 cagaagacac tttccaagct catcggacgt tctttgcgga cggtccaata cgcagtcaag
     7261 gacttggtgg ccgagcgctg gatctccgtc gtgaagctca acggccccgg caccgtgtcg
     7321 gcctacgtgg tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt gcgcctgtcg
     7381 gtgttcagtg ccgccgtggt ggttgatcac gacgaccagg acgaatcgct gttggggcat
     7441 ggcgacctgc gccgcatccc gaccctgtat ccgggcgagc agcaactacc gaccggcccc
     7501 ggcgaggagc cgcccagcca gcccggcatt ccgggcatgg aaccagacct gccagccttg
     7561 accgaaacgg aggaatggga acggcgcggg cagcagcgcc tgccgatgcc cgatgagccg
     7621 tgttttctgg acgatggcga gccgttggag ccgccgacac gggtcacgct gccgcgccgg
     7681 tagcacttgg gttgcgcagc aacccgtaag tgcgctgttc cagactatcg gctgtagccg
     7741 cctcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc
     7801 cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccgt ttttatcagg
     7861 ctctgggagg cagaataaat gatcatatcg tcaattatta cctccacggg gagagcctga
     7921 gcaaactggc
//
