LOCUS       sequence-177368-        3349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 transfer plasmid for pST44 polycistronic plasmid suite; 
            N-term cleavable single tag; contains DHFR gene in BamHI-NgoMIV 
            cassette as positive control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3349)
  AUTHORS   Tan S, Kern RC, Selleck W
  TITLE     The pST44 polycistronic expression system for producing protein 
            complexes in Escherichia coli.
  JOURNAL   Protein Expr Purif. 2005 Apr;40(2):385-95.
  PUBMED    15766881
REFERENCE   2  (bases 1 to 3349)
  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: "Protein 
            Expr Purif."; date: "2005-04"; volume: "40(2)"; pages: "385-95"
COMMENT     Sequence Label: pST50Trc2-HPCNDHFR
FEATURES             Location/Qualifiers
     source          1..3349
                     /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             137..172
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     CDS             194..214
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(781..799)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      795..842
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        907..998
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1863..1902
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1933..2521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2422..2441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2675..2692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2707..2846
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2772..2791)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2932..2954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2948..3139)
                     /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 atgtctgaag atcaggtaga tccacgttta atcgacggta aaggatctgg
      181 tggtggtggt ggtgaaaacc tgtacttcca gggatccatc agtctgattg cggcgctagc
      241 ggtagatcgc gttatcggca tggaaaacgc catgccatgg aacctgcctg ccgatctcgc
      301 ctggtttaaa cgcaacacct taaataaacc cgtgattatg gggcgccata cctgggaatc
      361 aatcggtagg cctttgcccg gccgcaaaaa tattatcctc agcagtcaac ccgggaccga
      421 tgatcgggtt acctgggtta aatcggtcga cgaagccatc gcggcgtgtg gtgacgtacc
      481 agaaatcatg gtgattggcg gcggacgcgt ttatgaacag ttcttgccaa aagcgcaaaa
      541 actgtatctg acgcatatcg atgcagaagt ggaaggcgac acccattttc cggattacga
      601 gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc agaactcgca
      661 tagctattgt ttcgaaatcc tcgagcgtcg tgccggctaa tgtacaaagc ttggtaccac
      721 gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      781 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      841 tgctgaaagg aggaactata tcgggattag cccgcctaat gagcgggctt ttttttaatt
      901 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      961 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1021 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1081 tcgtgaaagt aaaagacgca gaggaccaat tgggggcacg agtgggatac atagaactgg
     1141 acttgaatag cggtaaaatc cttgagagtt ttcgccctga agagcgtttt ccaatgatga
     1201 gcactttcaa agttctgcta tgtggagcag tattatcccg tgtagatgcg gggcaagagc
     1261 aactcggacg acgaatacac tattcgcaga atgacttggt tgaatactcc ccagtgacag
     1321 aaaagcacct tacggacgga atgacggtaa gagaattatg tagtgccgcc ataacgatga
     1381 gtgataacac tgcggcgaac ttacttctga caaccatcgg tggaccgaag gaattaaccg
     1441 cttttttgca caatatggga gaccatgtaa ctcgccttga ccgttgggaa ccagaactga
     1501 atgaagccat accaaacgac gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat
     1561 tacgcaaact attaactggc gaactactta ctctggcttc acggcaacaa ttaatagact
     1621 ggcttgaagc ggataaagtt gcaggaccac tactgcgttc ggcacttcct gctggctggt
     1681 ttattgctga taaatctggg gcaggagagc gtggttcacg gggtatcatt gccgcacttg
     1741 gaccagatgg taagccttcc cgtatcgtag ttatctacac gacgggtagt caggcaacta
     1801 tggacgaacg aaatagacag attgctgaaa taggggcttc actgattaag cattggtaaa
     1861 ccgatacaat taaaggctcc ttttggagcc tttttttttg gacggaccgg tagaaaagat
     1921 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1981 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2041 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2101 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2161 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2221 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2281 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2341 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2401 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2461 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2521 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2581 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2641 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2701 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     2761 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2821 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2881 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2941 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     3001 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3061 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3121 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atggggtaat
     3181 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     3241 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3301 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgtt
//
