LOCUS       sequence-413563-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Characterization plasmid for PPgk (sfgfp expressed from PPgk on the 
            S. aureus pC194 replicon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  AUTHORS   Rondthaler SN, Sarker B, Howitz N, Shah I, Andrews LB
  TITLE     Toolbox of Characterized Genetic Parts for Staphylococcus aureus.
  JOURNAL   ACS Synth Biol. 2023 Dec 8. doi: 10.1021/acssynbio.3c00325.
  PUBMED    38064657
REFERENCE   2  (bases 1 to 6572)
  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: "ACS Synth 
            Biol. 2023 Dec 8. doi: 10.1021/acssynbio.3c00325."
COMMENT     Sequence Label: pSR1057
FEATURES             Location/Qualifiers
     source          1..6572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1406..1423)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1406..1422)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1415..1437)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1631..1650)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1750..1772
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1810..1828)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1896..2000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2001..2069,2070..2861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2219..2238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3032..3620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3521..3540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3774..3791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3908..3929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3944..3949,3950..3967,3968..3974)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3982..3998
                     /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     3987..4009
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4006..4022
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4006..4022
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(4040..4057)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(4064..4774,4775..4777))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(4064..4777)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4104..4124)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4719..4747
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(5289..5303)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 aaaaaaacac aaaagaccac attttttaat gtggtctttt attcttcaac taaagcaccc
       61 attagttcaa caaacgaaaa ttggataaag tgggatattt ttaaaatata tatttatgtt
      121 acagtaatat tgacttttaa aaaaggattg attctaatga agaaagcaga caagtaagcc
      181 tcctaaattc actttagata aaaatttagg aggcatatca aatgaacttt aataaaattg
      241 atttagacaa ttggaagaga aaagagatat ttaatcatta tttgaaccaa caaacgactt
      301 ttagtataac cacagaaatt gatattagtg ttttataccg aaacataaaa caagaaggat
      361 ataaatttta ccctgcattt attttcttag tgacaagggt gataaactca aatacagctt
      421 ttagaactgg ttacaatagc gacggagagt taggttattg ggataagtta gagccacttt
      481 atacaatttt tgatggtgta tctaaaacat tctctggtat ttggactcct gtaaagaatg
      541 acttcaaaga gttttatgat ttataccttt ctgatgtaga gaaatataat ggttcgggga
      601 aattgtttcc caaaacacct atacctgaaa atgctttttc tctttctatt attccatgga
      661 cttcatttac tgggtttaac ttaaatatca ataataatag taattacctt ctacccatta
      721 ttacagcagg aaaattcatt aataaaggta attcaatata tttaccgcta tctttacagg
      781 tacatcattc tgtttgtgat ggttatcatg caggattgtt tatgaactct attcaggaat
      841 tgtcagatag gcctaatgac tggcttttat aatatgagat aatgccgact gtacttttta
      901 cagtcggttt tctaatgtca ctaacctgcc ccgttagttg aagaaggttt ttatattaca
      961 gctccagatc catatccttc tttttctgaa ccgacttctc ctttttcgct tctttattcc
     1021 aattgcttta ttgacgttga gcctcggaac ccttaacaat cccaaaactt gtcgaatggt
     1081 cggcttaata gctcacgcta tgccgacatt cgtctgcaag tttagttaag ggttcttctc
     1141 aacgcacaat aaattttctc ggcataaatg cgtggtctaa tttttatttt taataacctt
     1201 gatagcaaaa aatgccattc caatacaaaa ccacatacct ataatcgata accacataac
     1261 agtcataaaa ccactccttt ttaacaaact ttatcacaag aaatatttaa attttaaatg
     1321 cctttatttt gaattttaag gggcatttta aagatttagg ggtaaatcat atagttttat
     1381 gcctaaaaac ctacagaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1441 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1501 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1561 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     1621 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     1681 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     1741 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     1801 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1861 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2221 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2521 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     3181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3361 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3421 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3481 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3541 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3601 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3661 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3721 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3781 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3841 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3901 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     3961 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     4021 acatgattac gaattcttag tggtggtggt ggtggtggga tcctttgtag agctcatcca
     4081 tgccatgtgt aatcccagca gcagttacaa actcaagaag gaccatgtgg tcacgctttt
     4141 cgttaggatc tttcgaaagg acagattgtg tcgacaggta atggttgtct ggtaaaagga
     4201 cagggccatc gccaattgga gtattttgtt gataatggtc tgctagttga acggaaccat
     4261 cttcaacgtt gtggcgaatt ttgaagttag ctttgattcc attcttttgt ttgtctgccg
     4321 tgatgtatac attgtgtgag ttaaagttgt actcgagttt gtgtccaaga atgtttccat
     4381 cttctttaaa atcaataccc tttaactcga tacgattaac aagggtatca ccttcaaact
     4441 tgacttcagc acgcgtcttg taggtcccgt catctttgaa agatatagtg cgttcctgta
     4501 cataaccttc aggcatggca ctcttgaaaa agtcatgacg tttcatgtga tccggataac
     4561 gggaaaagca ttgaacacca taggtcagag tagtgacaag tgttggccac ggaacaggta
     4621 gttttccagt agtgcaaata aatttaaggg tgagttttcc gtttgtagca tcaccttcac
     4681 cctctccacg gacagaaaat ttgtgcccat taacatcacc atctaattca acaagaattg
     4741 ggacaactcc agtgaaaagt tcttctcctt tgctcataaa taatcatcct cctaaggtac
     4801 ccggggatcc gatgcatctt gctcgataca tttgcccgat aatatattga tatctaatct
     4861 ttatttatta tagatatgtt agtcataatt ttgcattaaa gcactccgct ttcattatag
     4921 cgcttctatt cgaaagaacc aagtcaggtc ctttctccca aattattcat tagacacagt
     4981 gattgtattt ctgggttagc atttggttta gtcaaaaata tcaggctagc ggatcctcta
     5041 gagtcgacct gcaggcatgc aagcttttaa aaagcaaata tgagccaaat aaatatattc
     5101 taattctaca aacaaaaatt tgagcaaatt cagtgtcgat tttttaagac actgcccagt
     5161 tacatgcaaa ttaaaatttt catgattttt tatagttcct aacagggtta aaatttgtat
     5221 aacgaaagta taatgtttat ataacgttag tataataaag cattttaaca ttatactttt
     5281 gataatcgtt tatcgtcgtc atcacaataa cttttaaaat actcgtgcat aattcaacag
     5341 ctgacctccc aataactaca tggtgttatc gggaggtcag ctgttagcac ttatattttg
     5401 ttattgttct tcctcgattt cgtctatcat tttgtgatta atttctcttt tttcttgttc
     5461 tgttaagtca taaagttcac tagctaaata ctctttttgt ttccaaatat aaaaaatttg
     5521 atagatatat tcggttggat caatttcttt taagtaatct aaatccccat tttttaattt
     5581 ctttttagcc tctttaaata atcctgaata aactaatacc tgtttacctt taagtgattt
     5641 ataaaatgca tcaaagactt tttgatttat taaataatca ctatctttac cagaatactt
     5701 agccatttca tataattctt tattattatt ttgtcttatt ttttgaactt gaacttgtgt
     5761 tatttctgaa atgcccgtta catcacgcca taaatctaac cattcttgtt ggctaatata
     5821 atatctttta tctgtgaaat acgatttatt tactgcaatt aacacatgaa aatgaggatt
     5881 ataatcatct ctttttttat tatatgtaat ctctaactta cgaacatatc cctttataac
     5941 actacctact ttttttctct ttataagttt tctaaaagaa ttattataac gttttatttc
     6001 attttctaat tcatcactca ttacattagg tgtagtcaaa gttaaaaaga taaactcctt
     6061 tttctcttgc tgcttaatat attgcatcat caaagataaa cccaatgcat cttttctagc
     6121 ttttctccaa gcacagacag gacaaaatcg atttttacaa gaattagctt tatataattt
     6181 ctgtttttct aaagttttat cagctacaaa agacagaaat gtattgcaat cttcaactaa
     6241 atccatttga ttctctccaa tatgacgttt aataaatttc tgaaatactt gatttctttg
     6301 ttttttctca gtatactttt ccatgttata acacataaaa acaacttagt tttcacaaac
     6361 tatgacaata aaaaaagttg ctttttcccc tttctatgta tgttttttac tagtcattta
     6421 aaacgataca ttaataggta cgaaaaagca actttttttg cgcttaaaac cagtcatacc
     6481 aataacttaa gggtaactag cctcgccggc aatagttacc cttattatca agataagaaa
     6541 gaaaaggatt tttcgctacg ctcaaatcct tt
//
