LOCUS       sequence-432476-        7975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid PaFtsH2NFtsH1 protease: PaftsH2SD 
            -PaftsH11-453-PaftsH2463-1881 in pJN105.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7975)
  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 7975)
  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: pSG075
FEATURES             Location/Qualifiers
     source          1..7975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(260..793)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(982..987,988..1004,1005..1010))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1280..1301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1316..1321,1322..1339,1340..1346)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1354..1370
                     /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     1359..1381
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1378..1394
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1378..1394
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1413..1433
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1415..1433
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1462..1478
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1463..1479
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1571..2449)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2476..2760
                     /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     2683..2702
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(4765..4784)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4766..4784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4794..4810)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4803..4825)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5519..6181)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      6182..6953
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 accttcggga gcgcctgaag cccgttctgg acgccctggg gccgttgaat cgggatatgc
       61 aggccaaggc cgccgcgatc atcaaggccg tgggcgaaaa gctgctgacg gaacagcggg
      121 aagtccagcg ccagaaacag gcccagcgcc agcaggaacg cgggcgcgca catttccccg
      181 aaaagtgcca cctgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
      241 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
      301 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
      361 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
      421 cggtggcaat gccctgcctc cggtgctctc cggagactgc gagatcatag atatagatct
      481 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
      541 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
      601 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
      661 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
      721 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
      781 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
      841 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
      901 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
      961 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt
     1021 gccttcatcc gtttccacgg tgtgcgtcca tgggcaaata ttatacgcaa ggcgacaagg
     1081 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     1141 gaatgcttaa tgaattacaa cagtttttat gcatgcgccc aatacgcaaa ccgcctctcc
     1201 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1261 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     1321 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1381 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     1441 agctgggtac cgggcccccc ctcgaggtcg acggtatcga tgcataatgt gcctgtcaaa
     1501 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     1561 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     1621 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     1681 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     1741 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     1801 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     1861 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     1921 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     1981 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     2041 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     2101 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     2161 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     2221 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     2281 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     2341 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     2401 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     2461 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
     2521 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     2581 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     2641 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     2701 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     2761 acccgttttt ttgggctagc ttgcgaggaa cattgcccat ggaaaagaaa gatcaatgga
     2821 acattggcta ctggatcgtc gccggcctgt tgctgctgac gctgcagaac tactggcagg
     2881 cggccaagac cgtcgagccc gtgccctaca gcgaattcga gaaggcgctg gccgaggggc
     2941 gcgtcgccga agtgctggtg tcggaccgca cggtcaccgg gcgcctgaaa tcgccggaca
     3001 gccggggcaa gaccaccatc gtggccactc gcgtcgaacc cgacctggcc gagcggctgt
     3061 ccaagtacga cgtgccttac gcgcgggtgg tggaaagcac ctggctacgt gatgtgctct
     3121 cctggattct gccggcggtg gccttcttcg gcgtctggtt cttcctgttc cgccgcttcg
     3181 ccgagaagca gggcatgggc ggcttcctga gcatcggcaa gagccgcgcc aaggtgttca
     3241 tggagaagaa caccggcgtc accttcgctg acgtcgccgg ctgcgacgag gccaaggagg
     3301 aggtcagcga gctggtcgag ttccttcgcg atccgggcaa gttccagcgc ctgggcggcc
     3361 gtatcccgcg cggcgtgctg atggtcggcc cgcccggcac cggcaagacc ctgctggcca
     3421 aggccatcgc cggcgaagcc aaggtgccgt tcttcaccat ttccggttcc gacttcgtcg
     3481 agatgttcgt cggcgtgggc gcttcccgtg tccgcgacat gttcgaccag gccaagaagc
     3541 atgcgccgtg catcatcttc atcgacgaga tcgacgccgt cggccgccat cgcggcgccg
     3601 gcctgggcgg tggtcacgac gagcgcgagc agaccctcaa ccagttgctg gtggagatgg
     3661 atggcttcga gatgaacgat ggcatcatcg tcatcgctgc caccaaccgt ccggacgttc
     3721 tggacccggc gctgctgcgt ccgggccgct tcgaccgcca ggtggtggtc ggcttgccgg
     3781 atatccgcgg gcgcgaacag atcctcaagg tgcacatgcg caaggtgccg ctgggcgacc
     3841 atgtcgatcc tgccgtcatc gcccgcggta cgcccggttt ctccggcgcc gacctggcca
     3901 acctggtcaa cgaggcctcg ctgttcgccg cgcgctccaa caagcgcatc gtcgacatgc
     3961 gcgagttcga gctggccaag gacaagatca tgatgggcgc cgagcgcaag accatggtca
     4021 tgtccgagaa ggagaagcgc aacaccgcct accacgaagc cggccacgcc atcgtcggtc
     4081 gcctggtgcc ggagcatgac ccggtctaca aggtgtccat cattccgcgc ggacgcgccc
     4141 tgggcgtgac catgttcctc cccgaggaag atcgctacag cctgtccaag cgcgcgctgg
     4201 aaagccagat ctgctcgctg ttcggcggcc gtatcgcgga agagatgacc ctcggcttcg
     4261 aaggcgtcac cactggcgct tccaacgaca tcatgcgtgc tacccagttg gcgcggaaca
     4321 tggtgaccaa gtggggtctg tccgagaagc tcggcccgct gatgtacgcg gaggaggagg
     4381 gcgaggtgtt cctcggccgt agcgccggca gccagcacgc caatgtgtcg ggcgagaccg
     4441 ccaagatgat cgaccaggaa gtccggcgca tcatcgacga ctgctacggt accgccaagc
     4501 gactgctcga cgagaaccgc gacaagctgg agatgatggc cgacgcactg atgaagtacg
     4561 aaaccatcga ctccgaccag atcgacgaca tcatggccgg tcgcgtgccg cgcgagccgc
     4621 gtgactggca gggtggttcg ggtaccggca ctccgccggc caacctggaa gagtcgggcc
     4681 gccgcgagaa cacccctccc atcggtgggc cggccggcga gcactgatct agagcggccg
     4741 ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4801 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4861 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4921 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     4981 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgta ctgcgatgag
     5041 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc
     5101 tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc
     5161 cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg
     5221 gtttattgac taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg
     5281 ctcaggctct ccccgtggag gtaataattg acgatatgat catttattct gcctcccaga
     5341 gcctgataaa aacggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt
     5401 ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata gggcggcgag
     5461 gcggctacag ccgatagtct ggaacagcgc acttacgggt tgctgcgcaa cccaagtgct
     5521 accggcgcgg cagcgtgacc cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac
     5581 acggctcatc gggcatcggc aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg
     5641 tcaaggctgg caggtctggt tccatgcccg gaatgccggg ctggctgggc ggctcctcgc
     5701 cggggccggt cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc
     5761 catgccccaa cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca
     5821 ccgacaggcg caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg
     5881 ccgacacggt gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt
     5941 ccttgactgc gtattggacc gtccgcaaag aacgtccgat gagcttggaa agtgtcttct
     6001 ggctgaccac cacggcgttc tggtggccca tctgcgccac gaggtgatgc agcagcattg
     6061 ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt
     6121 gcacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca
     6181 tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca
     6241 acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt
     6301 ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc
     6361 ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg
     6421 tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg
     6481 gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag
     6541 gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg
     6601 cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa
     6661 ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac
     6721 cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa
     6781 ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg
     6841 ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc
     6901 ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc
     6961 catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg
     7021 gagagcaagc ccgtagggcg ccgcagccgc cgtaggcggt cacgactttg cgaagcaaag
     7081 tctagtgagt atactcaagc attgagtggc ccgccggagg caccgccttg cgctgccccc
     7141 gtcgagccgg ttggacacca aaagggaggg gcaggcatgg cggcatacgc gatcatgcga
     7201 tgcaagaagc tggcgaaaat gggcaacgtg gcggccagtc tcaagcacgc ctaccgcgag
     7261 cgcgagacgc ccaacgctga cgccagcagg acgccagaga acgagcactg ggcggccagc
     7321 agcaccgatg aagcgatggg ccgactgcgc gagttgctgc cagagaagcg gcgcaaggac
     7381 gctgtgttgg cggtcgagta cgtcatgacg gccagcccgg aatggtggaa gtcggccagc
     7441 caagaacagc aggcggcgtt cttcgagaag gcgcacaagt ggctggcgga caagtacggg
     7501 gcggatcgca tcgtgacggc cagcatccac cgtgacgaaa ccagcccgca catgaccgcg
     7561 ttcgtggtgc cgctgacgca ggacggcagg ctgtcggcca aggagttcat cggcaacaaa
     7621 gcgcagatga cccgcgacca gaccacgttt gcggccgctg tggccgatct agggctgcaa
     7681 cggggcatcg agggcagcaa ggcacgtcac acgcgcattc aggcgttcta cgaggccctg
     7741 gagcggccac cagtgggcca cgtcaccatc agcccgcaag cggtcgagcc acgcgcctat
     7801 gcaccgcagg gattggccga aaagctggga atctcaaagc gcgttgagac gccggaagcc
     7861 gtggccgacc ggctgacaaa agcggttcgg caggggtatg agcctgccct acaggccgcc
     7921 gcaggagcgc gtgagatgcg caagaaggcc gatcaagccc aagagacggc ccgag
//
