LOCUS       sequence-340002-        9457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 250 kD Penn State ladder protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9457)
  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 9457)
  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: pPSU250
FEATURES             Location/Qualifiers
     source          1..9457
                     /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             2795..2818
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2828..2857
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3056..4153
                     /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      4010..4019
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4127..4150
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     primer_bind     complement(6889..6907)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6903..6950
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        7015..7106
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7971..8010
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      8041..8629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8530..8549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8783..8800
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8815..8954
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8880..8899)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9040..9062
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9056..9247)
                     /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 ctgctgataa
      421 caaatttaac aaagagcaac aaaacgcctt ttacgagatc ctgcatttac cgaacctgac
      481 cgaggaacag cgcaacggct ttattcagtc actgaaagac gacccgagtg tgagcaaaga
      541 aattctggca gaggccaaaa agctgaacga tgcccaagct ccgaaaggat ccggtagcag
      601 cggtagcagt ggttcaagcg caccggttcg tagcctgaat tgtaccctgc gtgatagcca
      661 gcagaaaagc ctggttatga gtggtccgta tgaactgaaa gcactgcatc tgcaaggtca
      721 ggatatggaa cagcaggttg tttttagcat gagctttgtt cagggtgaag aaagcaatga
      781 taaaattccg gttgcactgg gtctgaaaga gaaaaatctg tatctgagct gcgtgctgaa
      841 agatgataaa ccgaccttac agctggaaag cgttgatccg aaaaactacc cgaaaaagaa
      901 aatggaaaaa cgctttgtgt tcaacaaaat cgagatcaac aacaaacttg agtttgagag
      961 cgcacagttt ccgaattggt atattagcac cagccaggcc gaaaatatgc cggttttttt
     1021 aggtggcacc aaaggtggcc aggatattac cgattttacc atgcagtttg tgtccagcgc
     1081 cggcgaaggt cagggccagg gtcaaggtga aggttccggc agtgaggcag aagcccgccc
     1141 gactaacttt atccgtcaga tcatcgatga agatctggcc agtggtaagc acaccacagt
     1201 acacacccgt ttcccgccgg agccgaatgg ctatctgcat attggccatg cgaaatctat
     1261 ctgcctgaac ttcgggatcg cccaggacta taaaggccag tgcaacctgc gtttcgacga
     1321 cactaacccg gtaaaagaag atatcgagta tgttgagtcg atcaaaaacg acgtagagtg
     1381 gttaggtttt cactggtctg gtaacgtccg ttactcctcc gattattttg atcagctcca
     1441 cgcctatgcg atcgaactga tcaataaagg cctggcgtac gttgatgaac tgacgccgga
     1501 acagatccgc gaataccgcg gcaccctgac gcaaccgggt aaaaacagcc cgtaccgcga
     1561 ccgcagcgtt gaagagaacc tggcgctgtt cgaaaaaatg cgtgccggtg gttttgaaga
     1621 aggtaaagcc tgcctgcgtg cgaaaatcga catggcttca ccgtttatcg tgatgcgcga
     1681 tccggtgctg taccgtatta aatttgctga acaccaccag actggcaaca agtggtgcat
     1741 ctacccgatg tacgacttca cccactgcat cagcgatgcc ctggaaggta ttacgcactc
     1801 tctgtgtacg cttgagttcc aggacaaccg tcgtctgtac gactgggtac tggacaacat
     1861 cacgattcct gttcacccgc gccagtatga gttctcgcgc ctgaatctgg aatacaccgt
     1921 gatgtccaag cgtaagttga acctgctggt gaccgacaag cacgttgaag gctgggatga
     1981 cccgcgtatg ccgaccattt ccggtctgcg tcgtcgtggt tacactgcgg cttctattcg
     2041 tgagttctgc aaacgcatcg gcgtgaccaa gcaggacaac accattgaga tggcgtcgct
     2101 ggaatcctgc atccgtgaag atctcaacga aaatgcgccg cgcgcaatgg cggttatcga
     2161 tccggtgaaa ctggttatcg aaaactatca gggcgaaggc gaaatggtta ccatgccgaa
     2221 ccatccgaac aaaccggaaa tgggcagccg tcaggtgccg tttagcggtg agatttggat
     2281 tgatcgcgcc gatttccgcg aagaagctaa caagcagtac aaacgtctgg tgctgggtaa
     2341 agaagtgcgt ctgcgtaatg cttatgtgat taaggcagaa cgtgtcgaga aagatgccga
     2401 aggtaatatc accaccatct tctgtactta tgacgccgat accttaagca aagatccggc
     2461 agatggtcgt aaagtcaaag gtgttattca ctgggtgagc gcggcacatg cgctgccggt
     2521 tgaaatccgt ttgtatgacc gtctgttcag cgtgcctaac ccaggtgctg cggatgattt
     2581 cctgtcggtg attaacccgg aatcgctggt gatcaaacag ggctttgctg aaccgtcgct
     2641 gaaagatgcg gttgcgggta aagcattcca gtttgagcgt gaaggttact tctgcctcga
     2701 tagccgccat tctacggcgg aaaaaccggt atttaaccgc accgttgggc tgcgtgatac
     2761 ctgggcgaaa gtaggcgagc ggccgatggc aagctggtca catccgcagt ttgaaaaagg
     2821 tagcagtcat catcaccacc atcaccatca tcatcacagc agcggtagcg gtagcgcaga
     2881 taacaaattt aacaaagaac agcagaacgc cttctacgaa attctgcatc tgccgaatct
     2941 gaatgaagaa cagcgtaatg gttttatcca gagcctgaaa gatgatccga gccagagcgc
     3001 aaatctgctg gccgaagcaa aaaaactgaa tgatgcacag gcaccgaaag gatctaaaat
     3061 cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga
     3121 agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa
     3181 actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg
     3241 ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga
     3301 caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa
     3361 gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct
     3421 gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa
     3481 aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc
     3541 tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg
     3601 cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa
     3661 acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac
     3721 agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta
     3781 tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct
     3841 gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcgaaagagt tcctcgaaaa
     3901 ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt
     3961 agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga
     4021 aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc
     4081 cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa
     4141 agacgcgcag actaactctg gatccactca acagccacaa gccaaatacc gtcacgatta
     4201 tcgtgcgccg gattaccaga ttactgatat tgacttgacc tttgacctcg acgcgcaaaa
     4261 gacggtcgtt accgcggtca gccaggctgt ccgtcatggt gcatcagatg ctccccttcg
     4321 tctcaacggc gaagacctca aactggtttc tgttcatatt aatgatgagc cgtggaccgc
     4381 ctggaaagaa gaagagggcg cactggttat cagtaatttg ccggagcgtt ttacgcttaa
     4441 gatcattaat gaaataagcc cggcggcgaa taccgcgctg gaagggcttt atcagtcagg
     4501 cgatgcgctt tgcacccagt gtgaagccga aggtttccgc catattacgt attatctcga
     4561 ccgcccggac gtgctggcgc gttttaccac caaaattatt gccgataaaa tcaaatatcc
     4621 cttcctgctt tccaatggta accgcgttgc gcaaggcgaa ctggaaaacg gacgccattg
     4681 ggtacagtgg caggacccgt tcccgaaacc gtgctacctg tttgcgctgg tggcaggcga
     4741 ctttgatgta ctgcgcgata cctttaccac gcgttctggt cgcgaagtag cactggagct
     4801 gtacgtcgat cgcggcaacc ttgatcgcgc gccgtgggcg atgacctcgc tgaaaaactc
     4861 catgaaatgg gatgaagaac gctttggcct ggagtatgac ctcgacatct atatgatcgt
     4921 cgcggtggat ttcttcaata tgggcgcaat ggagaataag gggctgaata tctttaactc
     4981 caaatatgtg ctggcccgca ccgacaccgc caccgacaaa gattacctcg atattgaacg
     5041 cgttatcggc catgaatatt tccataactg gaccggtaac cgagtgacct gtcgcgactg
     5101 gttccagctc agcctgaaag aaggtttaac cgtcttccgc gatcaggagt tcagctctga
     5161 ccttggttcc cgcgcagtta accgcatcaa taatgtacgc accatgcgcg gattgcagtt
     5221 tgcagaagac gccagcccga tggcgcaccc gatccgcccg gatatggtca ttgagatgaa
     5281 caacttctac accctgaccg tttacgagaa gggcgcggaa gtgattcgca tgatccacac
     5341 cctgcttggc gaagaaaact tccagaaagg gatgcagctt tatttcgagc gtcatgatgg
     5401 tagtgcagcg acctgtgacg actttgtgca ggcgatggaa gatgcgtcga atgtcgatct
     5461 ctcccatttc cgccgttggt acagccagtc cggtacaccg attgtgaccg tcaaagacga
     5521 ctacaatccg gaaaccgagc agtacaccct gaccatcagc cagcgcacgc cagccacgcc
     5581 ggatcaggca gaaaaacagc cgctgcatat tccgtttgcc atcgaactgt atgataacga
     5641 aggcaaagtg atcccgttgc agaaaggcgg tcatccggtg aattccgtgc tgaacgtcac
     5701 tcaggcggaa cagacctttg tctttgataa tgtctacttc cagccggtgc ctgcgctgct
     5761 gtgcgaattc tctgcgccag tgaaactgga atataagtgg agcgatcagc aactgacctt
     5821 cctgatgcgt catgcgcgta atgatttctc ccgctgggat gcggcgcaaa gtttgctggc
     5881 aacctacatc aagctgaacg tcgcgcgtca tcagcaaggt cagccgctgt ctctgccggt
     5941 gcatgtggct gatgctttcc gcgcggtact gcttgatgag aagattgatc cagcgctggc
     6001 ggcagaaatc ctgacgctgc cttctgtcaa tgaaatggct gaattgttcg atatcatcga
     6061 cccgattgct attgccgaag tacgcgaagc actcactcgt actctggcga ctgaactggc
     6121 ggatgagcta ctggctattt acaacgcgaa ttaccagagc gagtaccgtg ttgagcatga
     6181 agatattgca aaacgcactc tgcgtaatgc ctgcctgcgt ttcctcgctt ttggtgaaac
     6241 gcatctggct gatgtgctgg tgagcaagca gttccacgaa gcaaacaata tgactgatgc
     6301 gctggcggcg ctttctgcgg cggttgccgc acagctgcct tgccgtgacg cgctgatgca
     6361 ggagtacgac gacaagtggc atcagaacgg tctggtgatg gataaatggt ttatcctgca
     6421 agccaccagc ccggcggcga atgtgctgga gacggtgcgc ggcctgttgc agcatcgctc
     6481 atttaccatg agcaacccga accgtattcg ttcgttgatt ggcgcgtttg cgggcagcaa
     6541 tccggcagcg ttccatgccg aagatggcag cggttacctg ttcctggtgg aaatgcttac
     6601 cgacctcaac agccgtaacc cgcaggtggc ttcacgtctg attgaaccgc tgattcgcct
     6661 gaaacgttac gatgccaaac gtcaggagaa aatgcgcgcg gcgctggaac agttgaaagg
     6721 gctggaaaat ctctctggcg atctgtacga gaagataact aaagcactgg ctcagtacac
     6781 taaaaacctg cagtaatgta caagatccag atctaagctt ggtaccacgc gtgcgcgctg
     6841 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     6901 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     6961 gaactatatc gggattagcc cgcctaatga gcgggctttt ttttaattcc cctatttgtt
     7021 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     7081 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7141 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctc gtgaaagtaa
     7201 aagacgcaga ggaccaattg ggggcacgag tgggatacat agaactggac ttgaatagcg
     7261 gtaaaatcct tgagagtttt cgccctgaag agcgttttcc aatgatgagc actttcaaag
     7321 ttctgctatg tggagcagta ttatcccgtg tagatgcggg gcaagagcaa ctcggacgac
     7381 gaatacacta ttcgcagaat gacttggttg aatactcccc agtgacagaa aagcacctta
     7441 cggacggaat gacggtaaga gaattatgta gtgccgccat aacgatgagt gataacactg
     7501 cggcgaactt acttctgaca accatcggtg gaccgaagga attaaccgct tttttgcaca
     7561 atatgggaga ccatgtaact cgccttgacc gttgggaacc agaactgaat gaagccatac
     7621 caaacgacga gcgagacacc acaatgcctg cggcaatggc aacaacatta cgcaaactat
     7681 taactggcga actacttact ctggcttcac ggcaacaatt aatagactgg cttgaagcgg
     7741 ataaagttgc aggaccacta ctgcgttcgg cacttcctgc tggctggttt attgctgata
     7801 aatctggggc aggagagcgt ggttcacggg gtatcattgc cgcacttgga ccagatggta
     7861 agccttcccg tatcgtagtt atctacacga cgggtagtca ggcaactatg gacgaacgaa
     7921 atagacagat tgctgaaata ggggcttcac tgattaagca ttggtaaacc gatacaatta
     7981 aaggctcctt ttggagcctt tttttttgga cggaccggta gaaaagatca aaggatcttc
     8041 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8101 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8161 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     8221 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8281 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8341 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8401 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8461 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8521 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8581 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8641 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     8701 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     8761 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     8821 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     8881 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     8941 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     9001 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     9061 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     9121 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     9181 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     9241 tggtcactga tgcctccgtg taagggggat ttctgttcat ggggtaatga taccgatgaa
     9301 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     9361 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     9421 tcaatgccag cgcttcgtta atacagatgt aggtgtt
//
