LOCUS       sequence-340001-        6808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 150 kD Penn State ladder protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6808)
  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 6808)
  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: pPSU150
FEATURES             Location/Qualifiers
     source          1..6808
                     /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(4240..4258)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4254..4301
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4366..4457
                     /gene="bla"
                     /label=AmpR promoter
     terminator      5322..5361
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      5392..5980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5881..5900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6134..6151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6166..6305
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6231..6250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6391..6413
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6407..6598)
                     /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 gttgagcatc 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 tataacaaag 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 aactctggat ccactcaaca gccacaagcc aaataccgtc acgattatcg
     1561 tgcgccggat taccagatta ctgatattga cttgaccttt gacctcgacg cgcaaaagac
     1621 ggtcgttacc gcggtcagcc aggctgtccg tcatggtgca tcagatgctc cccttcgtct
     1681 caacggcgaa gacctcaaac tggtttctgt tcatattaat gatgagccgt ggaccgcctg
     1741 gaaagaagaa gagggcgcac tggttatcag taatttgccg gagcgtttta cgcttaagat
     1801 cattaatgaa ataagcccgg cggcgaatac cgcgctggaa gggctttatc agtcaggcga
     1861 tgcgctttgc acccagtgtg aagccgaagg tttccgccat attacgtatt atctcgaccg
     1921 cccggacgtg ctggcgcgtt ttaccaccaa aattattgcc gataaaatca aatatccctt
     1981 cctgctttcc aatggtaacc gcgttgcgca aggcgaactg gaaaacggac gccattgggt
     2041 acagtggcag gacccgttcc cgaaaccgtg ctacctgttt gcgctggtgg caggcgactt
     2101 tgatgtactg cgcgatacct ttaccacgcg ttctggtcgc gaagtagcac tggagctgta
     2161 cgtcgatcgc ggcaaccttg atcgcgcgcc gtgggcgatg acctcgctga aaaactccat
     2221 gaaatgggat gaagaacgct ttggcctgga gtatgacctc gacatctata tgatcgtcgc
     2281 ggtggatttc ttcaatatgg gcgcaatgga gaataagggg ctgaatatct ttaactccaa
     2341 atatgtgctg gcccgcaccg acaccgccac cgacaaagat tacctcgata ttgaacgcgt
     2401 tatcggccat gaatatttcc ataactggac cggtaaccga gtgacctgtc gcgactggtt
     2461 ccagctcagc ctgaaagaag gtttaaccgt cttccgcgat caggagttca gctctgacct
     2521 tggttcccgc gcagttaacc gcatcaataa tgtacgcacc atgcgcggat tgcagtttgc
     2581 agaagacgcc agcccgatgg cgcacccgat ccgcccggat atggtcattg agatgaacaa
     2641 cttctacacc ctgaccgttt acgagaaggg cgcggaagtg attcgcatga tccacaccct
     2701 gcttggcgaa gaaaacttcc agaaagggat gcagctttat ttcgagcgtc atgatggtag
     2761 tgcagcgacc tgtgacgact ttgtgcaggc gatggaagat gcgtcgaatg tcgatctctc
     2821 ccatttccgc cgttggtaca gccagtccgg tacaccgatt gtgaccgtca aagacgacta
     2881 caatccggaa accgagcagt acaccctgac catcagccag cgcacgccag ccacgccgga
     2941 tcaggcagaa aaacagccgc tgcatattcc gtttgccatc gaactgtatg ataacgaagg
     3001 caaagtgatc ccgttgcaga aaggcggtca tccggtgaat tccgtgctga acgtcactca
     3061 ggcggaacag acctttgtct ttgataatgt ctacttccag ccggtgcctg cgctgctgtg
     3121 cgaattctct gcgccagtga aactggaata taagtggagc gatcagcaac tgaccttcct
     3181 gatgcgtcat gcgcgtaatg atttctcccg ctgggatgcg gcgcaaagtt tgctggcaac
     3241 ctacatcaag ctgaacgtcg cgcgtcatca gcaaggtcag ccgctgtctc tgccggtgca
     3301 tgtggctgat gctttccgcg cggtactgct tgatgagaag attgatccag cgctggcggc
     3361 agaaatcctg acgctgcctt ctgtcaatga aatggctgaa ttgttcgata tcatcgaccc
     3421 gattgctatt gccgaagtac gcgaagcact cactcgtact ctggcgactg aactggcgga
     3481 tgagctactg gctatttaca acgcgaatta ccagagcgag taccgtgttg agcatgaaga
     3541 tattgcaaaa cgcactctgc gtaatgcctg cctgcgtttc ctcgcttttg gtgaaacgca
     3601 tctggctgat gtgctggtga gcaagcagtt ccacgaagca aacaatatga ctgatgcgct
     3661 ggcggcgctt tctgcggcgg ttgccgcaca gctgccttgc cgtgacgcgc tgatgcagga
     3721 gtacgacgac aagtggcatc agaacggtct ggtgatggat aaatggttta tcctgcaagc
     3781 caccagcccg gcggcgaatg tgctggagac ggtgcgcggc ctgttgcagc atcgctcatt
     3841 taccatgagc aacccgaacc gtattcgttc gttgattggc gcgtttgcgg gcagcaatcc
     3901 ggcagcgttc catgccgaag atggcagcgg ttacctgttc ctggtggaaa tgcttaccga
     3961 cctcaacagc cgtaacccgc aggtggcttc acgtctgatt gaaccgctga ttcgcctgaa
     4021 acgttacgat gccaaacgtc aggagaaaat gcgcgcggcg ctggaacagt tgaaagggct
     4081 ggaaaatctc tctggcgatc tgtacgagaa gataactaaa gcactggctc agtacactaa
     4141 aaactaatgt acaagatcca gatctaagct tggtaccacg cgtgcgcgct gatccggctg
     4201 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     4261 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     4321 cgggattagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt ttatttttct
     4381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta aaagacgcag
     4561 aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc ggtaaaatcc
     4621 ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa gttctgctat
     4681 gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga cgaatacact
     4741 attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt acggacggaa
     4801 tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact gcggcgaact
     4861 tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac aatatgggag
     4921 accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata ccaaacgacg
     4981 agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta ttaactggcg
     5041 aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg gataaagttg
     5101 caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat aaatctgggg
     5161 caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt aagccttccc
     5221 gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga aatagacaga
     5281 ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt aaaggctcct
     5341 tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt cttgagatcc
     5401 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5461 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5521 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5581 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5641 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5701 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5761 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5821 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5881 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5941 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6001 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6061 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6121 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     6181 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     6241 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     6301 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     6361 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     6421 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     6481 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     6541 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     6601 atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga aacgagagag
     6661 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     6721 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     6781 gcgcttcgtt aatacagatg taggtgtt
//
