LOCUS       sequence-339998-        3568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 30 kD Penn State ladder protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3568)
  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 3568)
  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: pPSU30
FEATURES             Location/Qualifiers
     source          1..3568
                     /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             89..111
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             128..157
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             842..919
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     primer_bind     complement(1000..1018)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1014..1061
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1126..1217
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2082..2121
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2152..2740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2641..2660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2894..2911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2926..3065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2991..3010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3151..3173
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3167..3358)
                     /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 tccggagagc tcaataattt tgtttaactt taagaaggag atatacatat
      121 gggtagtcat catcaccacc atcaccatca tcatcatggt agcgcagata acaagttcaa
      181 taaagaacag cagaacgcct tctatgaaat tctgcatctg ccgaatctga atgaagaaca
      241 gcgtaatggt tttatccaga gcctgaaaga tgatccgagc cagagcgcaa atctgctggc
      301 cgaagcaaaa aaactgaatg atgcacaggc accgaaagga tccgctgata acaaatttaa
      361 caaagagcaa caaaacgcct tttacgagat cctgcattta ccgaacctga ccgaggaaca
      421 gcgcaacggc tttattcagt cactgaaaga cgacccgagt gtgagcaaag aaattctggc
      481 agaggccaaa aagctgaacg atgcccaagc tccggccgga ggtagcggta gcggttcagg
      541 ttctggttca ggtagtggta gcagcgaact ggaaaaagca atggttgcac tgattgatgt
      601 gtttcatcag tatagcggtc gtgaaggcga taaacacaaa ctgaaaaaaa gcgagctgaa
      661 agagctgatc aataacgaac tgagccattt tctggaagag atcaaagaac aagaggtggt
      721 ggataaagtt atggaaaccc tggataatga tggtgatggt gaatgcgatt ttcaagagtt
      781 tatggcattt gttgcaatgg ttaccaccgc ctgtcatgaa ttttttgaac acgaagccgg
      841 caagcgacga tggaaaaaga atttcatagc cgtctcagca gccaaccgct ttaagaaaat
      901 ctcatcctcc ggggcacttc tgtaatgtac aggtaccacg cgtgcgcgct gatccggctg
      961 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1021 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     1081 cgggattagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt ttatttttct
     1141 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1201 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1261 cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta aaagacgcag
     1321 aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc ggtaaaatcc
     1381 ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa gttctgctat
     1441 gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga cgaatacact
     1501 attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt acggacggaa
     1561 tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact gcggcgaact
     1621 tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac aatatgggag
     1681 accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata ccaaacgacg
     1741 agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta ttaactggcg
     1801 aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
     1861 caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
     1921 caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
     1981 gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
     2041 ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
     2101 tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt cttgagatcc
     2161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2281 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2941 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3001 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3061 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3121 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3181 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3241 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3301 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3361 atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag
     3421 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     3481 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     3541 gcgcttcgtt aatacagatg taggtgtt
//
