LOCUS       sequence-177373-        3289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 transfer plasmid for pST44 polycistronic plasmid suite; 
            C-term single tag; N-term non-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 3289)
  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 3289)
  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-DHFRHIS
FEATURES             Location/Qualifiers
     source          1..3289
                     /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             620..637
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(721..739)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      735..782
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        847..938
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1803..1842
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1873..2461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2362..2381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2615..2632
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2647..2786
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2712..2731)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2872..2894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2888..3079)
                     /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 atgggatcca tcagtctgat tgcggcgcta gcggtagatc gcgttatcgg
      181 catggaaaac gccatgccat ggaacctgcc tgccgatctc gcctggttta aacgcaacac
      241 cttaaataaa cccgtgatta tggggcgcca tacctgggaa tcaatcggta ggcctttgcc
      301 cggccgcaaa aatattatcc tcagcagtca acccgggacc gatgatcggg ttacctgggt
      361 taaatcggtc gacgaagcca tcgcggcgtg tggtgacgta ccagaaatca tggtgattgg
      421 cggcggacgc gtttatgaac agttcttgcc aaaagcgcaa aaactgtatc tgacgcatat
      481 cgatgcagaa gtggaaggcg acacccattt tccggattac gagccggatg actgggaatc
      541 ggtattcagc gaattccacg atgctgatgc gcagaactcg catagctatt gtttcgaaat
      601 cctcgagcgt cgtgccggcc accaccacca ccaccactaa tgtacaaagc ttggtaccac
      661 gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
      721 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      781 tgctgaaagg aggaactata tcgggattag cccgcctaat gagcgggctt ttttttaatt
      841 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      901 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      961 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1021 tcgtgaaagt aaaagacgca gaggaccaat tgggggcacg agtgggatac atagaactgg
     1081 acttgaatag cggtaaaatc cttgagagtt ttcgccctga agagcgtttt ccaatgatga
     1141 gcactttcaa agttctgcta tgtggagcag tattatcccg tgtagatgcg gggcaagagc
     1201 aactcggacg acgaatacac tattcgcaga atgacttggt tgaatactcc ccagtgacag
     1261 aaaagcacct tacggacgga atgacggtaa gagaattatg tagtgccgcc ataacgatga
     1321 gtgataacac tgcggcgaac ttacttctga caaccatcgg tggaccgaag gaattaaccg
     1381 cttttttgca caatatggga gaccatgtaa ctcgccttga ccgttgggaa ccagaactga
     1441 atgaagccat accaaacgac gagcgagaca ccacaatgcc tgcggcaatg gcaacaacat
     1501 tacgcaaact attaactggc gaactactta ctctggcttc acggcaacaa ttaatagact
     1561 ggcttgaagc ggataaagtt gcaggaccac tactgcgttc ggcacttcct gctggctggt
     1621 ttattgctga taaatctggg gcaggagagc gtggttcacg gggtatcatt gccgcacttg
     1681 gaccagatgg taagccttcc cgtatcgtag ttatctacac gacgggtagt caggcaacta
     1741 tggacgaacg aaatagacag attgctgaaa taggggcttc actgattaag cattggtaaa
     1801 ccgatacaat taaaggctcc ttttggagcc tttttttttg gacggaccgg tagaaaagat
     1861 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1921 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1981 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     2041 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2101 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2161 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2221 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2281 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2341 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2401 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2461 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2521 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2581 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2641 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     2701 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2761 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2821 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2881 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2941 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     3001 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     3061 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atggggtaat
     3121 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     3181 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3241 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgtt
//
