LOCUS       sequence-182529-        4607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Pseudomonas aeruginosa pqsE protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4607)
  AUTHORS   Oke M, Carter LG, Johnson KA, Liu H, McMahon SA, Yan X, Kerou M, 
            Weikart ND, Kadi N, Sheikh MA, Schmelz S, Dorward M, Zawadzki M, 
            Cozens C, Falconer H, Powers H, Overton IM, van Niekerk CA, Peng X, 
            Patel P, Garrett RA, Prangishvili D, Botting CH, Coote PJ, Dryden 
            DT, Barton GJ, Schwarz-Linek U, Challis GL, Taylor GL, White MF, 
            Naismith JH
  TITLE     The Scottish Structural Proteomics Facility: targets, methods and 
            outputs.
  JOURNAL   J Struct Funct Genomics. 2010 Jun;11(2):167-80. doi: 
            10.1007/s10969-010-9090-y. Epub 2010 Apr 24.
  PUBMED    20419351
REFERENCE   2  (bases 1 to 4607)
  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"; doi: 
            "10.1007/s10969-010-9090-y"; journalName: "J Struct Funct Genomics";
            date: "2010-06"; volume: "11"; issue: "2"; pages: "167-80"
COMMENT     Sequence Label: pEAHISPApqsE
FEATURES             Location/Qualifiers
     source          1..4607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        86..104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             136..158
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             178..195
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             217..237
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1193..1210
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1263..1281)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1277..1324
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1361..1816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1448..1467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1658..1679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1842..1946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1947..2015,2016..2807)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2165..2184)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2978..3566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3467..3486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3720..3737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3752..3891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3817..3836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3977..3999
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3993..4184)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
       61 gcccgagatc tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt
      121 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgtc gtactaccat
      181 caccatcacc atcacgatta cgacatccca acgaccgaaa acctgtattt tcagggcgtc
      241 atgatgaggc tttcggctcc cggtcaactg gatgatgacc tgtgcctgtt gggggacgtc
      301 caggtgccgg tgttcctgct gcgtctcggt gaggcgagct gggcgctggt tgaaggaggg
      361 atcagccggg atgccgaatt ggtttgggcg gacctgtgcc gctgggtcgc tgacccgtcc
      421 caggtgcact actggctgat cacccacaag cactacgacc actgcggcct gctgccctac
      481 ctgtgtccgc ggctgccgaa cgtacaggtc ctggcgtccg agcggacctg ccaggcctgg
      541 aagtcggaaa gcgcggtgcg ggtggtcgag cgcttgaacc ggcaactgtt gcgtgcggag
      601 cagcggttgc ccgaggcctg tgcctgggac gctctgccgg ttcgcgcggt ggccgacggc
      661 gagtggctgg agctgggacc gcggcatcgc ctgcaggtca tagaggccca cggccacagc
      721 gacgatcacg tggttttcta cgacgtgcga cgccgacgcc tgttctgcgg cgatgccctg
      781 ggcgagttcg acgaggcaga gggggtgtgg cggccgctgg tgttcgacga catggaggct
      841 tacctggagt ccctggaacg tctgcagcgt ctgccgaccc tgctgcaact gatcccggga
      901 cacggcggcc tgctgcgggg gcggctggcc gcggatgggg ccgagtcggc ctataccgag
      961 tgtctgcgcc tgtgccggcg gttgctctgg cgccagtcca tgggcgaatc cctcgacgaa
     1021 ctgagcgagg agctgcaccg cgcctggggt gggcagagcg tcgacttcct gcccggcgaa
     1081 ctgcacctgg ggagcatgcg ccggatgctg gagattctct cccgccaggc gctgcctctg
     1141 gactgaggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg agcaccacca
     1201 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1261 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1321 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1381 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1441 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1501 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1561 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1621 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1681 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1741 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1801 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1861 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1921 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1981 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2041 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2101 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2161 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2221 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2281 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2341 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2401 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2461 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2521 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2581 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2641 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2701 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2761 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2821 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2881 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2941 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3001 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3061 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3121 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3181 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3241 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3301 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3361 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3421 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3481 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3541 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3601 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3661 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3721 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3781 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3841 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3901 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3961 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4021 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4081 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4141 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4201 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4261 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4321 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4381 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4441 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4501 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4561 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcc
//
