LOCUS       sequence-432467-        7234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PaFtsH1-6xHis cloned between XbaI/NheI in pET-28a(+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7234)
  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 7234)
  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: pSG058
FEATURES             Location/Qualifiers
     source          1..7234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(205..234)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     CDS             complement(238..258)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    2212..2236
                     /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(2236..2255)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2237..2255)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2339..2358)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2564..2641
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2642..3724
                     /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(2661..2680)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4533..4724
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4718..4740)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4826..4965
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4881..4900
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4980..4997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5151..5739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5231..5250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5850..5869)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5861..6676
                     /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             5861..6676
                     /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(5934..5953)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6768..7223)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6905..6926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7117..7136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagctcagtg
      241 atgatgatga tgatggtgct cgccggccgg cccaccgatg ggaggggtgt tctcgcggcg
      301 gcccgactct tccaggttgg ccggcggagt gccggtaccc gaaccaccct gccagtcacg
      361 cggctcgcgc ggcacgcgac cggccatgat gtcgtcgatc tggtcggagt cgatggtttc
      421 gtacttcatc agtgcgtcgg ccatcatctc cagcttgtcg cggttctcgt cgagcagtcg
      481 cttggcggta ccgtagcagt cgtcgatgat gcgccggact tcctggtcga tcatcttggc
      541 ggtctcgccc gacacattgg cgtgctggct gccggcgcta cggccgagga acacctcgcc
      601 ctcctcctcc gcgtacatca gcgggccgag cttctcggac agaccccact tggtcaccat
      661 gttccgcgcc aactgggtag cacgcatgat gtcgttggaa gcgccagtgg tgacgccttc
      721 gaagccgagg gtcatctctt ccgcgatacg gccgccgaac agcgagcaga tctggctttc
      781 cagcgcgcgc ttggacaggc tgtagcgatc ttcctcgggg aggaacatgg tcacgcccag
      841 ggcgcgtccg cgcggaatga tggacacctt gtagaccggg tcatgctccg gcaccaggcg
      901 accgacgatg gcgtggccgg cttcgtggta ggcggtgttg cgcttctcct tctcggacat
      961 gaccatggtc ttgcgctcgg cgcccatcat gatcttgtcc ttggccagct cgaactcgcg
     1021 catgtcgacg atgcgcttgt tggagcgcgc ggcgaacagc gaggcctcgt tgaccaggtt
     1081 ggccaggtcg gcgccggaga aaccgggcgt accgcgggcg atgacggcag gatcgacatg
     1141 gtcgcccagc ggcaccttgc gcatgtgcac cttgaggatc tgttcgcgcc cgcggatatc
     1201 cggcaagccg accaccacct ggcggtcgaa gcggcccgga cgcagcagcg ccgggtccag
     1261 aacgtccgga cggttggtgg cagcgatgac gatgatgcca tcgttcatct cgaagccatc
     1321 catctccacc agcaactggt tgagggtctg ctcgcgctcg tcgtgaccac cgcccaggcc
     1381 ggcgccgcga tggcggccga cggcgtcgat ctcgtcgatg aagatgatgc acggcgcatg
     1441 cttcttggcc tggtcgaaca tgtcgcggac acgggaagcg cccacgccga cgaacatctc
     1501 gacgaagtcg gaaccggaaa tggtgaagaa cggcaccttg gcttcgccgg cgatggcctt
     1561 ggccagcagg gtcttgccgg tgccgggcgg gccgaccatc agcacgccgc gcgggatacg
     1621 gccgcccagg cgctggaact tgcccggatc gcgaaggaac tcgaccagct cgctgacctc
     1681 ctccttggcc tcgtcgcagc cggcgacgtc agcgaaggtg gtcttgacct ggtcctcgga
     1741 cagcaggcgc gccttgctct tgccgaagct catcgggccg ccgcgaccgc cgccgccacc
     1801 ctgcatctgg cgcatgaaga acatgaagac ggcgatgatc accaggatcg ggaagctggc
     1861 gaccagcaac tgggtccaga tgctctgctg ttcgggctgc ttgccctcga ccacgacgtt
     1921 gttgttgacc aggtcgccga tcaggccgtt gtcctggatc gccgggcgga tggtcttgaa
     1981 ggtatcgccg tcgctacgct tgccggtgat gacatagcca tccacggtca ctcgctcgac
     2041 cttgccatcc ttgacctgct ggatgaagtc cgagtagttc agcgtctgtg gctcgctcgg
     2101 gctcgagaaa ttgttcatca cggtcaccag taccgccgcg atgatcagcc acagaatcag
     2161 attctttgcc atgtcgttca attgactacc ctctgaagcc ggctctagag gggaattgtt
     2221 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc
     2281 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
     2341 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
     2401 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
     2461 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
     2521 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg
     2581 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
     2641 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     2701 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     2761 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     2821 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     2881 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     2941 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     3001 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     3061 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     3121 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     3181 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     3241 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     3301 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     3361 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     3421 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     3481 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     3541 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     3601 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     3661 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3721 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga
     3781 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3841 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3901 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     3961 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     4021 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga
     4081 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     4141 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     4201 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     4261 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     4321 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     4381 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     4441 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     4501 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     4561 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     4621 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     4681 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     4741 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     4801 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     4861 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg
     4921 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     4981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5341 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5581 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat
     5821 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     5881 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     5941 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     6001 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     6061 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     6121 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
     6181 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     6241 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     6301 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     6361 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     6421 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     6481 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     6541 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     6601 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     6661 cgatgagttt ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     6781 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     6841 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     6901 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     6961 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     7021 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7081 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7141 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     7201 taatgcgccg ctacagggcg cgtcccattc gcca
//
