LOCUS       sequence-177360-        3355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 2 transfer plasmid for pST44 polycistronic plasmid suite; 
            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 3355)
  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 3355)
  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-CBPDHFR
FEATURES             Location/Qualifiers
     source          1..3355
                     /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..214
                     /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"
     primer_bind     complement(787..805)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      801..848
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        913..1004
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1869..1908
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1939..2527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2428..2447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2681..2698
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2713..2852
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2778..2797)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2938..2960
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2954..3145)
                     /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 atgtctaagc gacgatggaa aaagaatttc atagccgtct cagcagccaa
      181 ccgctttaag aaaatctcat cctccggggc acttctggga tccatcagtc tgattgcggc
      241 gctagcggta gatcgcgtta tcggcatgga aaacgccatg ccatggaacc tgcctgccga
      301 tctcgcctgg tttaaacgca acaccttaaa taaacccgtg attatggggc gccatacctg
      361 ggaatcaatc ggtaggcctt tgcccggccg caaaaatatt atcctcagca gtcaacccgg
      421 gaccgatgat cgggttacct gggttaaatc ggtcgacgaa gccatcgcgg cgtgtggtga
      481 cgtaccagaa atcatggtga ttggcggcgg acgcgtttat gaacagttct tgccaaaagc
      541 gcaaaaactg tatctgacgc atatcgatgc agaagtggaa ggcgacaccc attttccgga
      601 ttacgagccg gatgactggg aatcggtatt cagcgaattc cacgatgctg atgcgcagaa
      661 ctcgcatagc tattgtttcg aaatcctcga gcgtcgtgcc ggctaatgta caaagcttgg
      721 taccacgcgt gcgcgctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      841 gttttttgct gaaaggagga actatatcgg gattagcccg cctaatgagc gggctttttt
      901 ttaattcccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      961 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1021 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1081 aaacgctcgt gaaagtaaaa gacgcagagg accaattggg ggcacgagtg ggatacatag
     1141 aactggactt gaatagcggt aaaatccttg agagttttcg ccctgaagag cgttttccaa
     1201 tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt atcccgtgta gatgcggggc
     1261 aagagcaact cggacgacga atacactatt cgcagaatga cttggttgaa tactccccag
     1321 tgacagaaaa gcaccttacg gacggaatga cggtaagaga attatgtagt gccgccataa
     1381 cgatgagtga taacactgcg gcgaacttac ttctgacaac catcggtgga ccgaaggaat
     1441 taaccgcttt tttgcacaat atgggagacc atgtaactcg ccttgaccgt tgggaaccag
     1501 aactgaatga agccatacca aacgacgagc gagacaccac aatgcctgcg gcaatggcaa
     1561 caacattacg caaactatta actggcgaac tacttactct ggcttcacgg caacaattaa
     1621 tagactggct tgaagcggat aaagttgcag gaccactact gcgttcggca cttcctgctg
     1681 gctggtttat tgctgataaa tctggggcag gagagcgtgg ttcacggggt atcattgccg
     1741 cacttggacc agatggtaag ccttcccgta tcgtagttat ctacacgacg ggtagtcagg
     1801 caactatgga cgaacgaaat agacagattg ctgaaatagg ggcttcactg attaagcatt
     1861 ggtaaaccga tacaattaaa ggctcctttt ggagcctttt tttttggacg gaccggtaga
     1921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2041 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2581 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2641 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2701 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2761 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     2821 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     2881 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2941 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     3001 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     3061 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     3121 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     3181 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     3241 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     3301 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgtt
//
