LOCUS       sequence-339999-        8438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpresses 20, 40, 60 and 80 kD Penn State ladder proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  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 8438)
  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: pPSU20-40-60-80
FEATURES             Location/Qualifiers
     source          1..8438
                     /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"
     primer_bind     668..685
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     669..685
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             698..720
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             737..766
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1130..1783
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     RBS             1827..1849
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1866..1889
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1899..1928
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             4102..4124
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4141..4170
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(5868..5886)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5882..5929
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5996..6087
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6952..6991
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      7022..7610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7511..7530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7764..7781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7796..7935
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7861..7880)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8021..8043
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8037..8228)
                     /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 ggatctgctg ataacaaatt taacaaagag caacaaaacg ccttttacga
      421 gatcctgcat ttaccgaacc tgaccgagga acagcgcaac ggctttattc agtcactgaa
      481 agacgacccg agtgtgagca aagaaattct ggcagaggcc aaaaagctga acgatgccca
      541 agctccgaaa ggatccggtg ccggccagga acagctgcaa cagaatttca tagccgtctc
      601 agcagccaac cgctttaaga aaatctcatc ctccggggca cttctgtaat gtacaagatc
      661 cagatcttgt aaaacgacgg ccagtgaatt caataatttt gtttaacttt aagaaggaga
      721 tatacatatg ggtagtcatc atcaccacca tcaccatcat catcatggta gcgcagataa
      781 caagttcaat aaagaacagc agaacgcctt ctatgaaatt ctgcatctgc cgaatctgaa
      841 tgaagaacag cgtaatggtt ttatccagag cctgaaagat gatccgagcc agagcgcaaa
      901 tctgctggcc gaagcaaaaa aactgaatga tgcacaggca ccgaaaggat ccgctgataa
      961 caaatttaac aaagagcaac aaaacgcctt ttacgagatc ctgcatttac cgaacctgac
     1021 cgaggaacag cgcaacggct ttattcagtc actgaaagac gacccgagtg tgagcaaaga
     1081 aattctggca gaggccaaaa agctgaacga tgcccaagct ccggccggct cccctatact
     1141 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
     1201 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
     1261 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
     1321 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
     1381 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
     1441 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
     1501 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
     1561 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
     1621 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
     1681 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
     1741 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa taatgtacaa agcttcgcca
     1801 gggttttccc agtcgagctc aataattttg tttaacttta agaaggagat atacatatgg
     1861 caagctggtc acatccgcag tttgaaaaag gtagcagtca tcatcaccac catcaccatc
     1921 atcatcacag cagcggtagc ggtagcgcag ataacaaatt taacaaagaa cagcagaacg
     1981 ccttctacga aattctgcat ctgccgaatc tgaatgaaga acagcgtaat ggttttatcc
     2041 agagcctgaa agatgatccg agccagagcg caaatctgct ggccgaagca aaaaaactga
     2101 atgatgcaca ggcaccgaaa ggatctgctg ataacaaatt taacaaagag caacaaaacg
     2161 ccttttacga gatcctgcat ttaccgaacc tgaccgagga acagcgcaac ggctttattc
     2221 agtcactgaa agacgacccg agtgtgagca aagaaattct ggcagaggcc aaaaagctga
     2281 acgatgccca agctccgaaa ggatccggta gcagcggtag cagtggttca agcgcaccgg
     2341 ttcgtagcct gaattgtacc ctgcgtgata gccagcagaa aagcctggtt atgagtggtc
     2401 cgtatgaact gaaagcactg catctgcaag gtcaggatat ggaacagcag gttgttttta
     2461 gcatgagctt tgttcagggt gaagaaagca atgataaaat tccggttgca ctgggtctga
     2521 aagagaaaaa tctgtatctg agctgcgtgc tgaaagatga taaaccgacc ttacagctgg
     2581 aaagcgttga tccgaaaaac tacccgaaaa agaaaatgga aaaacgcttt gtgttcaaca
     2641 aaatcgagat caacaacaaa cttgagtttg agagcgcaca gtttccgaat tggtatatta
     2701 gcaccagcca ggccgaaaat atgccggttt ttttaggtgg caccaaaggt ggccaggata
     2761 ttaccgattt taccatgcag tttgtgtcca gcgccggcag aaaggccctg actaataaca
     2821 acaatgccgg tgaagcggag cagcagccac cgaaggcgaa gcgcgccaga atcgcttttc
     2881 atctggagct gcccaagcgg aggaccgtgc tcggcaacgt gctagtctgc ggcaacggcg
     2941 atgtgggcca acttggtctc ggcgaggaca tactggagcg caagcgtctc agccctgtgg
     3001 cgggcatccc ggatgccgtg gacatatccg ccggtggcat gcacaacctg gtgctgacca
     3061 agtcagggga catttactca ttcggttgca atgacgaagg tgctctaggc cgtgacacca
     3121 gcgaggatgg cagcgagtca aagccggatc ttatcgattt gcccggcaag gctctgtgca
     3181 tttcggcggg agactcgcat tccgcttgtc tgctggagga tggccgtgtc ttcgcttggg
     3241 gctcgttccg ggactctcat ggcaacatgg gcctcaccat cgatggtaac aagcgtacgc
     3301 ccatcgatct gatggagggc actgtctgtt gtagcatcgc gtccggcgcc gatcatttgg
     3361 tcatcctgac cactgccgga aaggtcttta ccgtgggctg cgctgagcag ggacaactcg
     3421 ggcggctgtc ggagcgttcc atctccggag agggtcgtcg tggcaaacgc gacttgctgc
     3481 ggcccaccca gctcatcatc acgcgggcca aaccttttga ggccatctgg gccaccaact
     3541 actgcacatt catgcgcgag tcacagacac aggtcatctg ggccacaggg cttaacaact
     3601 tcaagcagct ggcccacgag acaaagggca aggagttcgc actgacgccc atcaagacgg
     3661 aactcaaaga tatccgacat attgccgggg gtcagcacca tactgtcatc ctgacaacgg
     3721 accttaagtg ttccgtagtg ggtcgtccgg agtacggtcg gcttggtctg ggcgatgtca
     3781 aggatgttgt ggagaagcca acgattgtaa agaagctgac cgaaaagatt gtgtccgttg
     3841 gatgtggcga agtctgttcc tatgccgtta caattgatgg caaactgtat tcctggggct
     3901 ccggtgtcaa caatcaacta ggcgttggcg acggcgatga tgaactcgaa ccgatcgtgg
     3961 ttgtcagcaa gaacacgcag gggaagcaca tgctgctcgc atctggcggt ggtcagcacg
     4021 caatcttttt ggtcaaagcg gacaaacagg accagtaatg tacaggtacc agcggataac
     4081 aatttcacat ccggaaataa ttttgtttaa ctttaagaag gagatataca tatgggtagt
     4141 catcatcacc accatcacca tcatcatcat ggtagcgcag ataacaagtt caataaagaa
     4201 cagcagaacg ccttctatga aattctgcat ctgccgaatc tgaatgaaga acagcgtaat
     4261 ggttttatcc agagcctgaa agatgatccg agccagagcg caaatctgct ggccgaagca
     4321 aaaaaactga atgatgcaca ggcaccgaaa ggatccgctg ataacaaatt taacaaagag
     4381 caacaaaacg ccttttacga gatcctgcat ttaccgaacc tgaccgagga acagcgcaac
     4441 ggctttattc agtcactgaa agacgacccg agtgtgagca aagaaattct ggcagaggcc
     4501 aaaaagctga acgatgccca agctccggcc ggaagaaagg ccctgactaa taacaacaat
     4561 gccggtgaag cggagcagca gccaccgaag gcgaagcgcg ccagaatcgc ttttcatctg
     4621 gagctgccca agcggaggac cgtgctcggc aacgtgctag tctgcggcaa cggcgatgtg
     4681 ggccaacttg gtctcggcga ggacatactg gagcgcaagc gtctcagccc tgtggcgggc
     4741 atcccggatg ccgtggacat atccgccggt ggcatgcaca acctggtgct gaccaagtca
     4801 ggggacattt actcattcgg ttgcaatgac gaaggtgctc taggccgtga caccagcgag
     4861 gatggcagcg agtcaaagcc ggatcttatc gatttgcccg gcaaggctct gtgcatttcg
     4921 gcgggagact cgcattccgc ttgtctgctg gaggatggcc gtgtcttcgc ttggggctcg
     4981 ttccgggact ctcatggcaa catgggcctc accatcgatg gtaacaagcg tacgcccatc
     5041 gatctgatgg agggcactgt ctgttgtagc atcgcgtccg gcgccgatca tttggtcatc
     5101 ctgaccactg ccggaaaggt ctttaccgtg ggctgcgctg agcagggaca actcgggcgg
     5161 ctgtcggagc gttccatctc cggagagggt cgtcgtggca aacgcgactt gctgcggccc
     5221 acccagctca tcatcacgcg ggccaaacct tttgaggcca tctgggccac caactactgc
     5281 acattcatgc gcgagtcaca gacacaggtc atctgggcca cagggcttaa caacttcaag
     5341 cagctggccc acgagacaaa gggcaaggag ttcgcactga cgcccatcaa gacggaactc
     5401 aaagatatcc gacatattgc cgggggtcag caccatactg tcatcctgac aacggacctt
     5461 aagtgttccg tagtgggtcg tccggagtac ggtcggcttg gtctgggcga tgtcaaggat
     5521 gttgtggaga agccaacgat tgtaaagaag ctgaccgaaa agattgtgtc cgttggatgt
     5581 ggcgaagtct gttcctatgc cgttacaatt gatggcaaac tgtattcctg gggctccggt
     5641 gtcaacaatc aactaggcgt tggcgacggc gatgatgaac tcgaaccgat cgtggttgtc
     5701 agcaagaaca cgcaggggaa gcacatgctg ctcgcatctg gcggtggtca gcacgcaatc
     5761 tttttggtca aagcggacaa acaggaccag gccggctaat gtacaacgcg tctcgagaga
     5821 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     5881 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     5941 aactatatcc gactagtagc ccgcctaatg agcgggcttt tttttaattc ccctatttgt
     6001 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6061 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6121 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct cgtgaaagta
     6181 aaagacgcag aggaccaatt gggggcacga gtgggataca tagaactgga cttgaatagc
     6241 ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc caatgatgag cactttcaaa
     6301 gttctgctat gtggagcagt attatcccgt gtagatgcgg ggcaagagca actcggacga
     6361 cgaatacact attcgcagaa tgacttggtt gaatactccc cagtgacaga aaagcacctt
     6421 acggacggaa tgacggtaag agaattatgt agtgccgcca taacgatgag tgataacact
     6481 gcggcgaact tacttctgac aaccatcggt ggaccgaagg aattaaccgc ttttttgcac
     6541 aatatgggag accatgtaac tcgccttgac cgttgggaac cagaactgaa tgaagccata
     6601 ccaaacgacg agcgagacac cacaatgcct gcggcaatgg caacaacatt acgcaaacta
     6661 ttaactggcg aactacttac tctggcttca cggcaacaat taatagactg gcttgaagcg
     6721 gataaagttg caggaccact actgcgttcg gcacttcctg ctggctggtt tattgctgat
     6781 aaatctgggg caggagagcg tggttcacgg ggtatcattg ccgcacttgg accagatggt
     6841 aagccttccc gtatcgtagt tatctacacg acgggtagtc aggcaactat ggacgaacga
     6901 aatagacaga ttgctgaaat aggggcttca ctgattaagc attggtaaac cgatacaatt
     6961 aaaggctcct tttggagcct ttttttttgg acggaccggt agaaaagatc aaaggatctt
     7021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7141 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     7801 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     7861 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     7921 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7981 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     8041 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     8101 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     8161 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     8221 ttggtcactg atgcctccgt gtaaggggga tttctgttca tggggtaatg ataccgatga
     8281 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     8341 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     8401 gtcaatgcca gcgcttcgtt aatacagatg taggtgtt
//
