LOCUS       sequence-433134-        7918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hybrid PaFtsH1NFtsH2 protease: PaftsH2SD 
            -PaftsH21-462-PaftsH1454-1920.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7918)
  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 7918)
  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: pSG071
FEATURES             Location/Qualifiers
     source          1..7918
                     /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(4708..4727)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4709..4727)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4737..4753)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4746..4768)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5462..6124)
                     /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      6125..6896
                     /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 ggcaaagaat ctgattctgt
     2821 ggctgatcat cgcggcggta ctggtgaccg tgatgaacaa tttctcgagc ccgagcgagc
     2881 cacagacgct gaactactcg gacttcatcc agcaggtcaa ggatggcaag gtcgagcgag
     2941 tgaccgtgga tggctatgtc atcaccggca agcgtagcga cggcgatacc ttcaagacca
     3001 tccgcccggc gatccaggac aacggcctga tcggcgacct ggtcaacaac aacgtcgtgg
     3061 tcgagggcaa gcagcccgaa cagcagagca tctggaccca gttgctggtc gccagcttcc
     3121 cgatcctggt gatcatcgcc gtcttcatgt tcttcatgcg ccagatgcag ggtggcggcg
     3181 gcggtcgcgg cggcccgatg agcttcggca agagcaaggc gcgcctgctg tccgaggacc
     3241 aggtcaagac cacctttgcc gatgtcgcgg gtgtcgatga ggccaaggcc gagctggtcg
     3301 aaatcgtcga cttcctgaag aacccgcagg attacggccg cctcggcgcg cgcatcccga
     3361 aaggtgtgtt gctggttggc ccgcccggca cgggcaagac cctgctggcc aaggccgtgg
     3421 cgggcgaggc cggggtaccg ttcttctcta tctccggctc ggagttcgtc gagatgtttg
     3481 tcggcgtggg cgcggcgcgg gtgcgcgacc tgttcgagca ggcgcgcggg caggcgccgg
     3541 ccatcatctt catcgatgaa ctcgacgcgc tgggccgtgc acgcggcgtc ggcgggccca
     3601 tcggcggcca cgacgagcgt gagcagacgc tcaaccagct gctcaccgag atggacggct
     3661 tcgacagctc ggtggggctg atcatcctcg ccgccaccaa ccgccccgaa atcctcgacc
     3721 aggcgctgct gcgtgccggc cgcttcgacc gccaggtgct ggtggaccgg cccgacaaga
     3781 agggacggct ggacatcctg aaagtccacg tcaagaaggt gacgctggct caggatatcg
     3841 atctcgaaca ggtggctgcg ctgaccacag gcttttcggg tgcagacctc gcgaacctgg
     3901 tcaacgaggc cgcgctggcc gcgacccggc gcaaagcgtc cgccgtggag ttgcaggatt
     3961 tcaccgccgc catcgagcgc atcgtggcgg gcctggagaa gaagaaccga gtgctcaatc
     4021 ccaaggagcg ggaaaccgtg gcctatcacg agatgggcca tgcgctggtg gcgctggcgc
     4081 tgcccggcac cgaccccgtg cacaagatct cgatcatccc gcgcggcatg ggcgcgctgg
     4141 gctacacgct gcaacgcccc accgaagacc gcttcctgat gacgcgcgcc gatctggaac
     4201 acaagatcgc cgtgctgctg ggcggtcgtg cggccgagaa gctcgtgttc ggcgagctgt
     4261 ccaccggggc tgccgatgac ctcgcacggg ccaccgacat tgcccgcgac atgatcaccc
     4321 gctttggcat ggacgaagga ctgggctaca tcgccttcga ggcgcagcgg ccccgctttc
     4381 tcgatacacc ggaactggcc cacggcggct gccgggtggc cgaatcgacc caggcgcgca
     4441 tcgatcaggc catccgcgac atcgtgatgg gcgtgttcga gcgcgcctac cggatcctcg
     4501 acaccaaccg cgcggtgctg gagcgttgtg cgcgcgaact gctggcgcgg gaaacgctcg
     4561 acgagaacga catccgccaa ctgacgcagg gcttgcagtc cgaatcccgc gcgtcggagc
     4621 ccctccagcc cgaaacctca ctctctacac cccaagggtc cacaccatga tctagagcgg
     4681 ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     4741 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4801 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4861 tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat attttgttaa
     4921 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gtactgcgat
     4981 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
     5041 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
     5101 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
     5161 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
     5221 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
     5281 agagcctgat aaaaacggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     5341 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     5401 gaggcggcta cagccgatag tctggaacag cgcacttacg ggttgctgcg caacccaagt
     5461 gctaccggcg cggcagcgtg acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa
     5521 aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt
     5581 cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg ggcggctcct
     5641 cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt
     5701 cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga
     5761 acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca ttgaccacgt
     5821 aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc tcggccacca
     5881 agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg gaaagtgtct
     5941 tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga tgcagcagca
     6001 ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg
     6061 tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg
     6121 ccatgcttat ctccatgcgg taggggtgcc gcacggttgc ggcaccatgc gcaatcagct
     6181 gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag tcaattacag
     6241 attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc tgttgcgtac
     6301 cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata tctagttctt
     6361 tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc ggctccccct
     6421 ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggccttc ggccttgccc
     6481 aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg gggggcaggc
     6541 gggcgggctt cgcccttcga ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt
     6601 caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc
     6661 aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag aaaattaaaa
     6721 aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg tggtcgaagg
     6781 ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc
     6841 agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgctcgc
     6901 ggccatcgtc cacatatcca cgggctggca agggagcgca gcgaccgcgc agggcgaagc
     6961 ccggagagca agcccgtagg gcgccgcagc cgccgtaggc ggtcacgact ttgcgaagca
     7021 aagtctagtg agtatactca agcattgagt ggcccgccgg aggcaccgcc ttgcgctgcc
     7081 cccgtcgagc cggttggaca ccaaaaggga ggggcaggca tggcggcata cgcgatcatg
     7141 cgatgcaaga agctggcgaa aatgggcaac gtggcggcca gtctcaagca cgcctaccgc
     7201 gagcgcgaga cgcccaacgc tgacgccagc aggacgccag agaacgagca ctgggcggcc
     7261 agcagcaccg atgaagcgat gggccgactg cgcgagttgc tgccagagaa gcggcgcaag
     7321 gacgctgtgt tggcggtcga gtacgtcatg acggccagcc cggaatggtg gaagtcggcc
     7381 agccaagaac agcaggcggc gttcttcgag aaggcgcaca agtggctggc ggacaagtac
     7441 ggggcggatc gcatcgtgac ggccagcatc caccgtgacg aaaccagccc gcacatgacc
     7501 gcgttcgtgg tgccgctgac gcaggacggc aggctgtcgg ccaaggagtt catcggcaac
     7561 aaagcgcaga tgacccgcga ccagaccacg tttgcggccg ctgtggccga tctagggctg
     7621 caacggggca tcgagggcag caaggcacgt cacacgcgca ttcaggcgtt ctacgaggcc
     7681 ctggagcggc caccagtggg ccacgtcacc atcagcccgc aagcggtcga gccacgcgcc
     7741 tatgcaccgc agggattggc cgaaaagctg ggaatctcaa agcgcgttga gacgccggaa
     7801 gccgtggccg accggctgac aaaagcggtt cggcaggggt atgagcctgc cctacaggcc
     7861 gccgcaggag cgcgtgagat gcgcaagaag gccgatcaag cccaagagac ggcccgag
//
