LOCUS       sequence-339971-        4447 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses 60 kD Penn State ladder protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4447)
  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 4447)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pPSU60
FEATURES             Location/Qualifiers
     source          1..4447
                     /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             101..123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             140..169
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1879..1897)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1893..1940
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2005..2096
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2961..3000
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      3031..3619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3520..3539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3773..3790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3805..3944
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3870..3889)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4030..4052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4046..4237)
                     /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 attcaataat tttgtttaac tttaagaagg
      121 agatatacat atgggtagtc atcatcacca ccatcaccat catcatcatg gtagcgcaga
      181 taacaagttc aataaagaac agcagaacgc cttctatgaa attctgcatc tgccgaatct
      241 gaatgaagaa cagcgtaatg gttttatcca gagcctgaaa gatgatccga gccagagcgc
      301 aaatctgctg gccgaagcaa aaaaactgaa tgatgcacag gcaccgaaag gatccgctga
      361 taacaaattt aacaaagagc aacaaaacgc cttttacgag atcctgcatt taccgaacct
      421 gaccgaggaa cagcgcaacg gctttattca gtcactgaaa gacgacccga gtgtgagcaa
      481 agaaattctg gcagaggcca aaaagctgaa cgatgcccaa gctccggccg gaagaaaggc
      541 cctgactaat aacaacaatg ccggtgaagc ggagcagcag ccaccgaagg cgaagcgcgc
      601 cagaatcgct tttcatctgg agctgcccaa gcggaggacc gtgctcggca acgtgctagt
      661 ctgcggcaac ggcgatgtgg gccaacttgg tctcggcgag gacatactgg agcgcaagcg
      721 tctcagccct gtggcgggca tcccggatgc cgtggacata tccgccggtg gcatgcacaa
      781 cctggtgctg accaagtcag gggacattta ctcattcggt tgcaatgacg aaggtgctct
      841 aggccgtgac accagcgagg atggcagcga gtcaaagccg gatcttatcg atttgcccgg
      901 caaggctctg tgcatttcgg cgggagactc gcattccgct tgtctgctgg aggatggccg
      961 tgtcttcgct tggggctcgt tccgggactc tcatggcaac atgggcctca ccatcgatgg
     1021 taacaagcgt acgcccatcg atctgatgga gggcactgtc tgttgtagca tcgcgtccgg
     1081 cgccgatcat ttggtcatcc tgaccactgc cggaaaggtc tttaccgtgg gctgcgctga
     1141 gcagggacaa ctcgggcggc tgtcggagcg ttccatctcc ggagagggtc gtcgtggcaa
     1201 acgcgacttg ctgcggccca cccagctcat catcacgcgg gccaaacctt ttgaggccat
     1261 ctgggccacc aactactgca cattcatgcg cgagtcacag acacaggtca tctgggccac
     1321 agggcttaac aacttcaagc agctggccca cgagacaaag ggcaaggagt tcgcactgac
     1381 gcccatcaag acggaactca aagatatccg acatattgcc gggggtcagc accatactgt
     1441 catcctgaca acggacctta agtgttccgt agtgggtcgt ccggagtacg gtcggcttgg
     1501 tctgggcgat gtcaaggatg ttgtggagaa gccaacgatt gtaaagaagc tgaccgaaaa
     1561 gattgtgtcc gttggatgtg gcgaagtctg ttcctatgcc gttacaattg atggcaaact
     1621 gtattcctgg ggctccggtg tcaacaatca actaggcgtt ggcgacggcg atgatgaact
     1681 cgaaccgatc gtggttgtca gcaagaacac gcaggggaag cacatgctgc tcgcatctgg
     1741 cggtggtcag cacgcaatct ttttggtcaa agcggacaaa caggaccagg ccggctaatg
     1801 tacaaagctt ggtaccacgc gtgcgcgctg atccggctgc taacaaagcc cgaaaggaag
     1861 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1921 gggtcttgag gggttttttg ctgaaaggag gaactatatc gggattagcc cgcctaatga
     1981 gcgggctttt ttttaattcc cctatttgtt tatttttcta aatacattca aatatgtatc
     2041 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2101 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2161 ttgctcaccc agaaacgctc gtgaaagtaa aagacgcaga ggaccaattg ggggcacgag
     2221 tgggatacat agaactggac ttgaatagcg gtaaaatcct tgagagtttt cgccctgaag
     2281 agcgttttcc aatgatgagc actttcaaag ttctgctatg tggagcagta ttatcccgtg
     2341 tagatgcggg gcaagagcaa ctcggacgac gaatacacta ttcgcagaat gacttggttg
     2401 aatactcccc agtgacagaa aagcacctta cggacggaat gacggtaaga gaattatgta
     2461 gtgccgccat aacgatgagt gataacactg cggcgaactt acttctgaca accatcggtg
     2521 gaccgaagga attaaccgct tttttgcaca atatgggaga ccatgtaact cgccttgacc
     2581 gttgggaacc agaactgaat gaagccatac caaacgacga gcgagacacc acaatgcctg
     2641 cggcaatggc aacaacatta cgcaaactat taactggcga actacttact ctggcttcac
     2701 ggcaacaatt aatagactgg cttgaagcgg ataaagttgc aggaccacta ctgcgttcgg
     2761 cacttcctgc tggctggttt attgctgata aatctggggc aggagagcgt ggttcacggg
     2821 gtatcattgc cgcacttgga ccagatggta agccttcccg tatcgtagtt atctacacga
     2881 cgggtagtca ggcaactatg gacgaacgaa atagacagat tgctgaaata ggggcttcac
     2941 tgattaagca ttggtaaacc gatacaatta aaggctcctt ttggagcctt tttttttgga
     3001 cggaccggta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3061 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3721 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3781 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3841 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3901 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3961 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4021 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4081 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4141 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4201 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     4261 ttctgttcat ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     4321 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     4381 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     4441 aggtgtt
//
