LOCUS       sequence-339996-        8069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpresses 10, 30, 50 and 100 kD Penn State ladder proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  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 8069)
  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: pPSU10-30-50-100
FEATURES             Location/Qualifiers
     source          1..8069
                     /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             77..99
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             116..139
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             149..178
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             377..1474
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1331..1340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1448..1471
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     primer_bind     1499..1516
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1500..1516
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1529..1551
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1568..1591
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1601..1630
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             4242..4264
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4281..4310
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             4995..5072
                     /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"
     RBS             5122..5144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5161..5184
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             5194..5223
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(5499..5517)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5513..5560
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5627..5718
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6583..6622
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      6653..7241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7142..7161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7395..7412
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7427..7566
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7492..7511)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7652..7674
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7668..7859)
                     /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 tccctctaga aataattttg tttaacttta agaaggagat atacatatgg caagctggtc
      121 acatccgcag tttgaaaaag gtagcagtca tcatcaccac catcaccatc atcatcacag
      181 cagcggtagc ggtagcgcag ataacaaatt taacaaagaa cagcagaacg ccttctacga
      241 aattctgcat ctgccgaatc tgaatgaaga acagcgtaat ggttttatcc agagcctgaa
      301 agatgatccg agccagagcg caaatctgct ggccgaagca aaaaaactga atgatgcaca
      361 ggcaccgaaa ggatctaaaa tcgaagaagg taaactggta atctggatta acggcgataa
      421 aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt
      481 caccgttgag cacccggata aactggaaga gaaattccca caggttgcgg caactggcga
      541 tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct
      601 gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga
      661 tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct
      721 gatttataac aaggatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct
      781 ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta
      841 cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa
      901 gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct
      961 ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc
     1021 tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat
     1081 cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc
     1141 caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct
     1201 ggcgaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa
     1261 agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc
     1321 acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca
     1381 gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca
     1441 gactgtcgat gaagccctga aagacgcgca gactaactaa tgtacaagat ccagatcttg
     1501 taaaacgacg gccagtgaat tcaataattt tgtttaactt taagaaggag atatacatat
     1561 ggcaagctgg tcacatccgc agtttgaaaa aggtagcagt catcatcacc accatcacca
     1621 tcatcatcac agcagcggta gcggtagcgc agataacaaa tttaacaaag aacagcagaa
     1681 cgccttctac gaaattctgc atctgccgaa tctgaatgaa gaacagcgta atggttttat
     1741 ccagagcctg aaagatgatc cgagccagag cgcaaatctg ctggccgaag caaaaaaact
     1801 gaatgatgca caggcaccga aaggatctgc tgataacaaa tttaacaaag agcaacaaaa
     1861 cgccttttac gagatcctgc atttaccgaa cctgaccgag gaacagcgca acggctttat
     1921 tcagtcactg aaagacgacc cgagtgtgag caaagaaatt ctggcagagg ccaaaaagct
     1981 gaacgatgcc caagctccga aaggatccgg tagcagcggt agcagtggtt caagcgcacc
     2041 ggttcgtagc ctgaattgta ccctgcgtga tagccagcag aaaagcctgg ttatgagtgg
     2101 tccgtatgaa ctgaaagcac tgcatctgca aggtcaggat atggaacagc aggttgtttt
     2161 tagcatgagc tttgttcagg gtgaagaaag caatgataaa attccggttg cactgggtct
     2221 gaaagagaaa aatctgtatc tgagctgcgt gctgaaagat gataaaccga ccttacagct
     2281 ggaaagcgtt gatccgaaaa actacccgaa aaagaaaatg gaaaaacgct ttgtgttcaa
     2341 caaaatcgag atcaacaaca aacttgagtt tgagagcgca cagtttccga attggtatat
     2401 tagcaccagc caggccgaaa atatgccggt ttttttaggt ggcaccaaag gtggccagga
     2461 tattaccgat tttaccatgc agtttgtgtc cagcgccggc gaaggtcagg gccagggtca
     2521 aggtgaaggt tccggcagtg aggcagaagc ccgcccgact aactttatcc gtcagatcat
     2581 cgatgaagat ctggccagtg gtaagcacac cacagtacac acccgtttcc cgccggagcc
     2641 gaatggctat ctgcatattg gccatgcgaa atctatctgc ctgaacttcg ggatcgccca
     2701 ggactataaa ggccagtgca acctgcgttt cgacgacact aacccggtaa aagaagatat
     2761 cgagtatgtt gagtcgatca aaaacgacgt agagtggtta ggttttcact ggtctggtaa
     2821 cgtccgttac tcctccgatt attttgatca gctccacgcc tatgcgatcg aactgatcaa
     2881 taaaggcctg gcgtacgttg atgaactgac gccggaacag atccgcgaat accgcggcac
     2941 cctgacgcaa ccgggtaaaa acagcccgta ccgcgaccgc agcgttgaag agaacctggc
     3001 gctgttcgaa aaaatgcgtg ccggtggttt tgaagaaggt aaagcctgcc tgcgtgcgaa
     3061 aatcgacatg gcttcaccgt ttatcgtgat gcgcgatccg gtgctgtacc gtattaaatt
     3121 tgctgaacac caccagactg gcaacaagtg gtgcatctac ccgatgtacg acttcaccca
     3181 ctgcatcagc gatgccctgg aaggtattac gcactctctg tgtacgcttg agttccagga
     3241 caaccgtcgt ctgtacgact gggtactgga caacatcacg attcctgttc acccgcgcca
     3301 gtatgagttc tcgcgcctga atctggaata caccgtgatg tccaagcgta agttgaacct
     3361 gctggtgacc gacaagcacg ttgaaggctg ggatgacccg cgtatgccga ccatttccgg
     3421 tctgcgtcgt cgtggttaca ctgcggcttc tattcgtgag ttctgcaaac gcatcggcgt
     3481 gaccaagcag gacaacacca ttgagatggc gtcgctggaa tcctgcatcc gtgaagatct
     3541 caacgaaaat gcgccgcgcg caatggcggt tatcgatccg gtgaaactgg ttatcgaaaa
     3601 ctatcagggc gaaggcgaaa tggttaccat gccgaaccat ccgaacaaac cggaaatggg
     3661 cagccgtcag gtgccgttta gcggtgagat ttggattgat cgcgccgatt tccgcgaaga
     3721 agctaacaag cagtacaaac gtctggtgct gggtaaagaa gtgcgtctgc gtaatgctta
     3781 tgtgattaag gcagaacgtg tcgagaaaga tgccgaaggt aatatcacca ccatcttctg
     3841 tacttatgac gccgatacct taagcaaaga tccggcagat ggtcgtaaag tcaaaggtgt
     3901 tattcactgg gtgagcgcgg cacatgcgct gccggttgaa atccgtttgt atgaccgtct
     3961 gttcagcgtg cctaacccag gtgctgcgga tgatttcctg tcggtgatta acccggaatc
     4021 gctggtgatc aaacagggct ttgctgaacc gtcgctgaaa gatgcggttg cgggtaaagc
     4081 attccagttt gagcgtgaag gttacttctg cctcgatagc cgccattcta cggcggaaaa
     4141 accggtattt aaccgcaccg ttgggctgcg tgatacctgg gcgaaagtag gcgagtaatg
     4201 tacaaagctt cgccagggtt ttcccagtcg agctcaataa ttttgtttaa ctttaagaag
     4261 gagatataca tatgggtagt catcatcacc accatcacca tcatcatcat ggtagcgcag
     4321 ataacaagtt caataaagaa cagcagaacg ccttctatga aattctgcat ctgccgaatc
     4381 tgaatgaaga acagcgtaat ggttttatcc agagcctgaa agatgatccg agccagagcg
     4441 caaatctgct ggccgaagca aaaaaactga atgatgcaca ggcaccgaaa ggatccgctg
     4501 ataacaaatt taacaaagag caacaaaacg ccttttacga gatcctgcat ttaccgaacc
     4561 tgaccgagga acagcgcaac ggctttattc agtcactgaa agacgacccg agtgtgagca
     4621 aagaaattct ggcagaggcc aaaaagctga acgatgccca agctccggcc ggaggtagcg
     4681 gtagcggttc aggttctggt tcaggtagtg gtagcagcga actggaaaaa gcaatggttg
     4741 cactgattga tgtgtttcat cagtatagcg gtcgtgaagg cgataaacac aaactgaaaa
     4801 aaagcgagct gaaagagctg atcaataacg aactgagcca ttttctggaa gagatcaaag
     4861 aacaagaggt ggtggataaa gttatggaaa ccctggataa tgatggtgat ggtgaatgcg
     4921 attttcaaga gtttatggca tttgttgcaa tggttaccac cgcctgtcat gaattttttg
     4981 aacacgaagc cggcaagcga cgatggaaaa agaatttcat agccgtctca gcagccaacc
     5041 gctttaagaa aatctcatcc tccggggcac ttctgtaatg tacaggtacc agcggataac
     5101 aatttcacat ccggaaataa ttttgtttaa ctttaagaag gagatataca tatggcaagc
     5161 tggtcacatc cgcagtttga aaaaggtagc agtcatcatc accaccatca ccatcatcat
     5221 cacagcagcg gtagcggtag cgcagataac aaatttaaca aagaacagca gaacgccttc
     5281 tacgaaattc tgcatctgcc gaatctgaat gaagaacagc gtaatggttt tatccagagc
     5341 ctgaaagatg atccgagcca gagcgcaaat ctgctggccg aagcaaaaaa actgaatgat
     5401 gcacaggcac cgaaaggatc ctaagccggc tgtacaacgc gtctcgagag atccggctgc
     5461 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     5521 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     5581 cgactagtag cccgcctaat gagcgggctt ttttttaatt cccctatttg tttatttttc
     5641 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5701 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5761 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tcgtgaaagt aaaagacgca
     5821 gaggaccaat tgggggcacg agtgggatac atagaactgg acttgaatag cggtaaaatc
     5881 cttgagagtt ttcgccctga agagcgtttt ccaatgatga gcactttcaa agttctgcta
     5941 tgtggagcag tattatcccg tgtagatgcg gggcaagagc aactcggacg acgaatacac
     6001 tattcgcaga atgacttggt tgaatactcc ccagtgacag aaaagcacct tacggacgga
     6061 atgacggtaa gagaattatg tagtgccgcc ataacgatga gtgataacac tgcggcgaac
     6121 ttacttctga caaccatcgg tggaccgaag gaattaaccg cttttttgca caatatggga
     6181 gaccatgtaa ctcgccttga ccgttgggaa ccagaactga atgaagccat accaaacgac
     6241 gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat tacgcaaact attaactggc
     6301 gaactactta ctctggcttc acggcaacaa ttaatagact ggcttgaagc ggataaagtt
     6361 gcaggaccac tactgcgttc ggcacttcct gctggctggt ttattgctga taaatctggg
     6421 gcaggagagc gtggttcacg gggtatcatt gccgcacttg gaccagatgg taagccttcc
     6481 cgtatcgtag ttatctacac gacgggtagt caggcaacta tggacgaacg aaatagacag
     6541 attgctgaaa taggggcttc actgattaag cattggtaaa ccgatacaat taaaggctcc
     6601 ttttggagcc tttttttttg gacggaccgg tagaaaagat caaaggatct tcttgagatc
     6661 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6721 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6781 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6841 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6901 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6961 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7021 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7081 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7141 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7201 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7261 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7321 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7381 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     7441 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     7501 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     7561 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     7621 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     7681 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     7741 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     7801 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     7861 gatgcctccg tgtaaggggg atttctgttc atggggtaat gataccgatg aaacgagaga
     7921 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     7981 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     8041 agcgcttcgt taatacagat gtaggtgtt
//
