LOCUS       sequence-340003-        4177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 50 kD Penn State ladder protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4177)
  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 4177)
  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: pPSU50
FEATURES             Location/Qualifiers
     source          1..4177
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             152..175
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             185..214
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             413..1510
                     /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      1367..1376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1484..1507
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     primer_bind     complement(1609..1627)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1623..1670
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1735..1826
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2691..2730
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2761..3349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3250..3269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3503..3520
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3535..3674
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3600..3619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3760..3782
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3776..3967)
                     /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 tccaattgga attcgctagc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatggcaag ctggtcacat ccgcagtttg aaaaaggtag
      181 cagtcatcat caccaccatc accatcatca tcacagcagc ggtagcggta gcgcagataa
      241 caaatttaac aaagaacagc agaacgcctt ctacgaaatt ctgcatctgc cgaatctgaa
      301 tgaagaacag cgtaatggtt ttatccagag cctgaaagat gatccgagcc agagcgcaaa
      361 tctgctggcc gaagcaaaaa aactgaatga tgcacaggca ccgaaaggat ctaaaatcga
      421 agaaggtaaa ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt
      481 cggtaagaaa ttcgagaaag ataccggaat taaagtcacc gttgagcacc cggataaact
      541 ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc
      601 acacgaccgc tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa
      661 agcgttccag gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct
      721 gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt tataacaagg atctgctgcc
      781 gaacccgcca aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg
      841 taagagcgcg ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc
      901 tgacgggggt tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt
      961 ggataacgct ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca
     1021 catgaatgca gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc
     1081 gatgaccatc aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg
     1141 tgtaacggta ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag
     1201 cgcaggtatt aacgccgcca gtccgaacaa agagctggcg aaagagttcc tcgaaaacta
     1261 tctgctgact gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc
     1321 gctgaagtct tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa
     1381 cgcccagaaa ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt
     1441 gcgtactgcg gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga
     1501 cgcgcagact aactaatgta caagatccag atctaagctt ggtaccacgc gtgcgcgctg
     1561 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     1621 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1681 gaactatatc gggattagcc cgcctaatga gcgggctttt ttttaattcc cctatttgtt
     1741 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1801 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1861 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctc gtgaaagtaa
     1921 aagacgcaga ggaccaattg ggggcacgag tgggatacat agaactggac ttgaatagcg
     1981 gtaaaatcct tgagagtttt cgccctgaag agcgttttcc aatgatgagc actttcaaag
     2041 ttctgctatg tggagcagta ttatcccgtg tagatgcggg gcaagagcaa ctcggacgac
     2101 gaatacacta ttcgcagaat gacttggttg aatactcccc agtgacagaa aagcacctta
     2161 cggacggaat gacggtaaga gaattatgta gtgccgccat aacgatgagt gataacactg
     2221 cggcgaactt acttctgaca accatcggtg gaccgaagga attaaccgct tttttgcaca
     2281 atatgggaga ccatgtaact cgccttgacc gttgggaacc agaactgaat gaagccatac
     2341 caaacgacga gcgagacacc acaatgcctg cggcaatggc aacaacatta cgcaaactat
     2401 taactggcga actacttact ctggcttcac ggcaacaatt aatagactgg cttgaagcgg
     2461 ataaagttgc aggaccacta ctgcgttcgg cacttcctgc tggctggttt attgctgata
     2521 aatctggggc aggagagcgt ggttcacggg gtatcattgc cgcacttgga ccagatggta
     2581 agccttcccg tatcgtagtt atctacacga cgggtagtca ggcaactatg gacgaacgaa
     2641 atagacagat tgctgaaata ggggcttcac tgattaagca ttggtaaacc gatacaatta
     2701 aaggctcctt ttggagcctt tttttttgga cggaccggta gaaaagatca aaggatcttc
     2761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2881 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3001 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3061 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3421 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3481 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3541 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     3601 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3661 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3721 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3781 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3841 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3901 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3961 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     4021 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     4081 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     4141 tcaatgccag cgcttcgtta atacagatgt aggtgtt
//
