LOCUS       sequence-177375-        3295 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Position 2 transfer plasmid for pST44 polycistronic plasmid suite; 
            C-term 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 3295)
  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 3295)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-DHFRFLAG
FEATURES             Location/Qualifiers
     source          1..3295
                     /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..643
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(727..745)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      741..788
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        853..944
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1809..1848
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1879..2467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2368..2387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2621..2638
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2653..2792
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2718..2737)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2878..2900
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2894..3085)
                     /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 cgtgccggcg actacaagga cgatgatgac aagtaatgta caaagcttgg
      661 taccacgcgt gcgcgctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      721 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      781 gttttttgct gaaaggagga actatatcgg gattagcccg cctaatgagc gggctttttt
      841 ttaattcccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      901 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      961 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1021 aaacgctcgt gaaagtaaaa gacgcagagg accaattggg ggcacgagtg ggatacatag
     1081 aactggactt gaatagcggt aaaatccttg agagttttcg ccctgaagag cgttttccaa
     1141 tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt atcccgtgta gatgcggggc
     1201 aagagcaact cggacgacga atacactatt cgcagaatga cttggttgaa tactccccag
     1261 tgacagaaaa gcaccttacg gacggaatga cggtaagaga attatgtagt gccgccataa
     1321 cgatgagtga taacactgcg gcgaacttac ttctgacaac catcggtgga ccgaaggaat
     1381 taaccgcttt tttgcacaat atgggagacc atgtaactcg ccttgaccgt tgggaaccag
     1441 aactgaatga agccatacca aacgacgagc gagacaccac aatgcctgcg gcaatggcaa
     1501 caacattacg caaactatta actggcgaac tacttactct ggcttcacgg caacaattaa
     1561 tagactggct tgaagcggat aaagttgcag gaccactact gcgttcggca cttcctgctg
     1621 gctggtttat tgctgataaa tctggggcag gagagcgtgg ttcacggggt atcattgccg
     1681 cacttggacc agatggtaag ccttcccgta tcgtagttat ctacacgacg ggtagtcagg
     1741 caactatgga cgaacgaaat agacagattg ctgaaatagg ggcttcactg attaagcatt
     1801 ggtaaaccga tacaattaaa ggctcctttt ggagcctttt tttttggacg gaccggtaga
     1861 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1921 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1981 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2041 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2101 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2161 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2221 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2281 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2341 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2401 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2461 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2521 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2581 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2641 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2701 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     2761 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     2821 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2881 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     2941 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3001 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3061 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     3121 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3181 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3241 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgtt
//
