LOCUS       sequence-339994-        3292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 20 kD Penn State ladder protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3292)
  AUTHORS   Santilli RT, Williamson JE 3rd, Shibata Y, Sowers RP, Fleischman AN,
            Tan S
  TITLE     The Penn State Protein Ladder system for inexpensive protein 
            molecular weight markers.
  JOURNAL   Sci Rep. 2021 Aug 18;11(1):16703. doi: 10.1038/s41598-021-96051-x.
  PUBMED    34408191
REFERENCE   2  (bases 1 to 3292)
  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: "Sci Rep."; 
            date: "2021-08-18"; volume: "11(1):16703. doi"; pages: " 
            10.1038/s41598-021-96051-x"
COMMENT     Sequence Label: pPSU20
FEATURES             Location/Qualifiers
     source          1..3292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             83..105
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             122..145
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             155..184
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(724..742)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      738..785
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        850..941
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1806..1845
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1876..2464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2365..2384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2618..2635
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2650..2789
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2715..2734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2875..2897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2891..3082)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggcaccatgc atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
       61 tccctccggc tccggaaata attttgttta actttaagaa ggagatatac atatggcaag
      121 ctggtcacat ccgcagtttg aaaaaggtag cagtcatcat caccaccatc accatcatca
      181 tcacagcagc ggtagcggta gcgcagataa caaatttaac aaagaacagc agaacgcctt
      241 ctacgaaatt ctgcatctgc cgaatctgaa tgaagaacag cgtaatggtt ttatccagag
      301 cctgaaagat gatccgagcc agagcgcaaa tctgctggcc gaagcaaaaa aactgaatga
      361 tgcacaggca ccgaaaggat ctgctgataa caaatttaac aaagagcaac aaaacgcctt
      421 ttacgagatc ctgcatttac cgaacctgac cgaggaacag cgcaacggct ttattcagtc
      481 actgaaagac gacccgagtg tgagcaaaga aattctggca gaggccaaaa agctgaacga
      541 tgcccaagct ccgaaaggat ccggtgccgg ccaggaacag ctgcaacaga atttcatagc
      601 cgtctcagca gccaaccgct ttaagaaaat ctcatcctcc ggggcacttc tgtaatgtac
      661 aacgcgtgcg cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      721 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      781 ttttgctgaa aggaggaact atatcgggat tagcccgcct aatgagcggg ctttttttta
      841 attcccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      901 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      961 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1021 cgctcgtgaa agtaaaagac gcagaggacc aattgggggc acgagtggga tacatagaac
     1081 tggacttgaa tagcggtaaa atccttgaga gttttcgccc tgaagagcgt tttccaatga
     1141 tgagcacttt caaagttctg ctatgtggag cagtattatc ccgtgtagat gcggggcaag
     1201 agcaactcgg acgacgaata cactattcgc agaatgactt ggttgaatac tccccagtga
     1261 cagaaaagca ccttacggac ggaatgacgg taagagaatt atgtagtgcc gccataacga
     1321 tgagtgataa cactgcggcg aacttacttc tgacaaccat cggtggaccg aaggaattaa
     1381 ccgctttttt gcacaatatg ggagaccatg taactcgcct tgaccgttgg gaaccagaac
     1441 tgaatgaagc cataccaaac gacgagcgag acaccacaat gcctgcggca atggcaacaa
     1501 cattacgcaa actattaact ggcgaactac ttactctggc ttcacggcaa caattaatag
     1561 actggcttga agcggataaa gttgcaggac cactactgcg ttcggcactt cctgctggct
     1621 ggtttattgc tgataaatct ggggcaggag agcgtggttc acggggtatc attgccgcac
     1681 ttggaccaga tggtaagcct tcccgtatcg tagttatcta cacgacgggt agtcaggcaa
     1741 ctatggacga acgaaataga cagattgctg aaataggggc ttcactgatt aagcattggt
     1801 aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacggac cggtagaaaa
     1861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1981 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2581 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2641 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     2701 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     2761 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     2821 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     2881 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     2941 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     3001 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     3061 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggt
     3121 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3181 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3241 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg tt
//
