LOCUS       sequence-310313-        2925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-term 2xStrepII uncleavable Amp resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2925)
  AUTHORS   Altmannova V, Blaha A, Astrinidis S, Reichle H, Weir JR
  TITLE     InteBac - An integrated bacterial and baculovirus expression vector 
            suite.
  JOURNAL   Protein Sci. 2020 Sep 21. doi: 10.1002/pro.3957.
  PUBMED    32955754
REFERENCE   2  (bases 1 to 2925)
  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 
            Sci. 2020 Sep 21. doi: 10.1002/pro.3957."
COMMENT     Sequence Label: 6 pCOLI_A_C_STREP
FEATURES             Location/Qualifiers
     source          1..2925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        47..65
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             103..125
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(194..199,200..223,224..259,260..283)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(377..395)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      391..438
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        503..594
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1459..1498
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1529..2117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2018..2037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2271..2288
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2303..2442
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2368..2387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2528..2550
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2544..2735)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ttaatacaga tgtaggtgtt ggcaccatgc atcgatcccg cgaaattaat acgactcact
       61 atagggagac cacaacggtt tccctccggc tccggaaata attttgttta actttaagaa
      121 ggagactgga tccgaatcta gagcctgcag tctcgacaag cttggaagcg gatctggatc
      181 tggaagcagc gccagcgctt ggagtcatcc ccaatttgag aaaggtggag gttccggagg
      241 tggatcggga ggttcggcgt ggagccaccc gcagttcgaa aaataataag tcgagaagta
      301 ctagaggatg tacaacgcgt gcgcgctgat ccggctgcta acaaagcccg aaaggaagct
      361 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
      421 gtcttgaggg gttttttgct gaaaggagga actatatcgg gattagcccg cctaatgagc
      481 gggctttttt ttaattcccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      541 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      601 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      661 gctcacccag aaacgctcgt gaaagtaaaa gacgcagagg accaattggg ggcacgagtg
      721 ggatacatag aactggactt gaatagcggt aaaatccttg agagttttcg ccctgaagag
      781 cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gagcagtatt atcccgtgta
      841 gatgcggggc aagagcaact cggacgacga atacactatt cgcagaatga cttggttgaa
      901 tactccccag tgacagaaaa gcaccttacg gacggaatga cggtaagaga attatgtagt
      961 gccgccataa cgatgagtga taacactgcg gcgaacttac ttctgacaac catcggtgga
     1021 ccgaaggaat taaccgcttt tttgcacaat atgggagacc atgtaactcg ccttgaccgt
     1081 tgggaaccag aactgaatga agccatacca aacgacgagc gagacaccac aatgcctgcg
     1141 gcaatggcaa caacattacg caaactatta actggcgaac tacttactct ggcttcacgg
     1201 caacaattaa tagactggct tgaagcggat aaagttgcag gaccactact gcgttcggca
     1261 cttcctgctg gctggtttat tgctgataaa tctggggcag gagagcgtgg ttcacggggt
     1321 atcattgccg cacttggacc agatggtaag ccttcccgta tcgtagttat ctacacgacg
     1381 ggtagtcagg caactatgga cgaacgaaat agacagattg ctgaaatagg ggcttcactg
     1441 attaagcatt ggtaaaccga tacaattaaa ggctcctttt ggagcctttt tttttggacg
     1501 gaccggtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1561 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1621 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     1681 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1741 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1801 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1861 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1921 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1981 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2041 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2101 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2161 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2221 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2281 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2341 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2401 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     2461 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     2521 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     2581 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     2641 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     2701 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     2761 ctgttcatgg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     2821 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     2881 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcg
//
