LOCUS       sequence-177347-        3418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 1 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 3418)
  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 3418)
  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: pST50Trc1-CBPNDHFR
FEATURES             Location/Qualifiers
     source          1..3418
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..226
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             251..271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(850..868)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      864..911
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        976..1067
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1932..1971
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2002..2590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2491..2510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2744..2761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2776..2915
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2841..2860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3001..3023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3017..3208)
                     /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 attcgctagc tctagaaata attttgttta
      121 actttaagaa ggagatatac atatgtctaa gcgacgatgg aaaaagaatt tcatagccgt
      181 ctcagcagcc aaccgcttta agaaaatctc atcctccggg gcacttctgg gatctggtgg
      241 tggtggtggt gaaaacctgt acttccaggg atccatcagt ctgattgcgg cgctagcggt
      301 agatcgcgtt atcggcatgg aaaacgccat gccatggaac ctgcctgccg atctcgcctg
      361 gtttaaacgc aacaccttaa ataaacccgt gattatgggg cgccatacct gggaatcaat
      421 cggtaggcct ttgcccggcc gcaaaaatat tatcctcagc agtcaacccg ggaccgatga
      481 tcgggttacc tgggttaaat cggtcgacga agccatcgcg gcgtgtggtg acgtaccaga
      541 aatcatggtg attggcggcg gacgcgttta tgaacagttc ttgccaaaag cgcaaaaact
      601 gtatctgacg catatcgatg cagaagtgga aggcgacacc cattttccgg attacgagcc
      661 ggatgactgg gaatcggtat tcagcgaatt ccacgatgct gatgcgcaga actcgcatag
      721 ctattgtttc gaaatcctcg agcgtcgtgc cggctaatgt acaagatcca gatctaagct
      781 tggtaccacg cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
      841 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
      901 ggggtttttt gctgaaagga ggaactatat cgggattagc ccgcctaatg agcgggcttt
      961 tttttaattc ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1021 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1081 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1141 cagaaacgct cgtgaaagta aaagacgcag aggaccaatt gggggcacga gtgggataca
     1201 tagaactgga cttgaatagc ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc
     1261 caatgatgag cactttcaaa gttctgctat gtggagcagt attatcccgt gtagatgcgg
     1321 ggcaagagca actcggacga cgaatacact attcgcagaa tgacttggtt gaatactccc
     1381 cagtgacaga aaagcacctt acggacggaa tgacggtaag agaattatgt agtgccgcca
     1441 taacgatgag tgataacact gcggcgaact tacttctgac aaccatcggt ggaccgaagg
     1501 aattaaccgc ttttttgcac aatatgggag accatgtaac tcgccttgac cgttgggaac
     1561 cagaactgaa tgaagccata ccaaacgacg agcgagacac cacaatgcct gcggcaatgg
     1621 caacaacatt acgcaaacta ttaactggcg aactacttac tctggcttca cggcaacaat
     1681 taatagactg gcttgaagcg gataaagttg caggaccact actgcgttcg gcacttcctg
     1741 ctggctggtt tattgctgat aaatctgggg caggagagcg tggttcacgg ggtatcattg
     1801 ccgcacttgg accagatggt aagccttccc gtatcgtagt tatctacacg acgggtagtc
     1861 aggcaactat ggacgaacga aatagacaga ttgctgaaat aggggcttca ctgattaagc
     1921 attggtaaac cgatacaatt aaaggctcct tttggagcct ttttttttgg acggaccggt
     1981 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2041 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2101 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2161 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2221 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2281 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2341 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2401 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2461 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2521 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2581 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2641 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2701 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2761 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2821 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     2881 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     2941 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3001 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3061 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3121 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3181 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3241 tggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3301 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3361 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgtt
//
