LOCUS       sequence-190115-        5176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Pseudomonas aeruginosa orf PA5006 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5176)
  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 5176)
  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: pEAHISPA5006
FEATURES             Location/Qualifiers
     source          1..5176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        493..597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(598..666,667..1458)
                     /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(816..835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1629..2217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2118..2137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2371..2388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2403..2542
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2468..2487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2628..2650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2644..2835)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3344..3363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3344..3362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3394..3416
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3436..3453
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3475..3495
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5020..5037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5090..5108)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5104..5151
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     2461 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     2521 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     2581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     2701 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     2761 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     2821 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     2881 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     2941 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3001 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3061 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3121 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3181 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3241 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3301 tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata
     3361 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
     3421 catatgtcgt actaccatca ccatcaccat cacgattacg acatcccaac gaccgaaaac
     3481 ctgtattttc agggcgccat gagactggcc gaactgcaag ccgccgggcg ccagccgcaa
     3541 ctgccattgc gggtgaccct ggcggatggc gccggcgctg ccgaactgcg cctggacagt
     3601 ctgctgcggg tgctgcccgg gcagcgctac gtcggtgccg gcgaatggcg tgggcgcaag
     3661 gtgctggcca agctgctggt gggcggcaag gccgcacggc atttccagcg cgaactggaa
     3721 ggggcgcgtc tgctcgcggc ccaggggctg accaccccgg cgctgctggc cgacgggctg
     3781 cagcagggcg agggcggctg gttgctgttc gactacctcg acgcggcgca gagcctcggt
     3841 gacgcctggc gtgcggtgga ggcccaggcg ccgctgggcg atgcccagca agcggtgctc
     3901 ggcgaagccc tggcagccat tggcgagctg catgcaaagg gcctctggca ggacgacctg
     3961 cacctggaca acctgttgcg ccacgccggc cgcctgtatc tcatcgacgg tggcggcgtg
     4021 cgtgcggaaa ctcccggcca gccgctgtcg cgacagaaag tgctggaaaa cctcggggtg
     4081 ttcttcgccc agttgccggc gctcctcgat cctttcctgg aggagttgct ggtgcattac
     4141 ctgctggcca acggcgagca tgcgctgccg ctggaagcct tgcagaagga aatcgccaag
     4201 gttcgccgct ggcgggtgcg cgacctgctg agaaaggtcg gtcgcgactg tagcctgttc
     4261 agcgtcgtgc gcggtccgtt ccgcctgttc gccgcccgcc gcagcgaggc cgagacgatg
     4321 gcgcccctgc tggcggacgc ggaccgcctg atcgccgccg gccacgtcta caaggacggc
     4381 ggcgcggcca ccgtcgcccg ggtcgaagcg aacggccgca cgctgatcat caagcgctac
     4441 aacatcaagg gctgggtcca ttggctgaag cgcttctggc ggccgagccg ggcctgggcc
     4501 tcctggcgcg aaggcaaccg cctggaattc ctcggcatcg ccaccgcccg tccgctggcg
     4561 atgctggagc ggcgctggtg ctggctgcgc gggcgcgcct acctgattac cgattatctc
     4621 ggcggtcagg atataatcgc tcgttttaag ccttgcctgg acatgcccga tggcgccgtt
     4681 tcgccaccgg aagccgagtt ggcggcactg gatcggctgt tttcgacgtt gatccgcgaa
     4741 cgcatcagcc acggcgacct gaagggacac aacctgttct ggcaggccca tgaacaaggc
     4801 ggcgaatggg cgctgatcga cctggacgcg atgcgcaagc atcacagcca ggcgtccttt
     4861 gcccgcgcct acgcccgcga ccgcgcgcgc ttcctgcgca actggccgca ggacagcggc
     4921 ctctaccggc tgctggagca acggattccc cgcctcggcg acgccgccgg gtaggatccg
     4981 aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccactga
     5041 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     5101 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     5161 ggaactatat ccggat
//
