LOCUS       sequence-318773-        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     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             106..128
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(197..202,203..226,227..262,263..286)
                     /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(380..398)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      394..441
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        506..597
                     /gene="bla"
                     /label=AmpR promoter
     terminator      1462..1501
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1532..2120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2021..2040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2274..2291
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2306..2445
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2371..2390)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2531..2553
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2547..2738)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcgttaatac agatgtaggt gttggcacca tgcatcgatc ccgcgaaatt aatacgactc
       61 actataggga gaccacaacg gtttccctcc ggctccggaa ataattttgt ttaactttaa
      121 gaaggagact ggatccgaat ctagagcctg cagtctcgac aagcttggaa gcggatctgg
      181 atctggaagc agcgccagcg cttggagtca tccccaattt gagaaaggtg gaggttccgg
      241 aggtggatcg ggaggttcgg cgtggagcca cccgcagttc gaaaaataat aagtcgagaa
      301 gtactagagg atgtacaacg cgtgcgcgct gatccggctg ctaacaaagc ccgaaaggaa
      361 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      421 cgggtcttga ggggtttttt gctgaaagga ggaactatat cgggattagc ccgcctaatg
      481 agcgggcttt tttttaattc ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct cgtgaaagta aaagacgcag aggaccaatt gggggcacga
      721 gtgggataca tagaactgga cttgaatagc ggtaaaatcc ttgagagttt tcgccctgaa
      781 gagcgttttc caatgatgag cactttcaaa gttctgctat gtggagcagt attatcccgt
      841 gtagatgcgg ggcaagagca actcggacga cgaatacact attcgcagaa tgacttggtt
      901 gaatactccc cagtgacaga aaagcacctt acggacggaa tgacggtaag agaattatgt
      961 agtgccgcca taacgatgag tgataacact gcggcgaact tacttctgac aaccatcggt
     1021 ggaccgaagg aattaaccgc ttttttgcac aatatgggag accatgtaac tcgccttgac
     1081 cgttgggaac cagaactgaa tgaagccata ccaaacgacg agcgagacac cacaatgcct
     1141 gcggcaatgg caacaacatt acgcaaacta ttaactggcg aactacttac tctggcttca
     1201 cggcaacaat taatagactg gcttgaagcg gataaagttg caggaccact actgcgttcg
     1261 gcacttcctg ctggctggtt tattgctgat aaatctgggg caggagagcg tggttcacgg
     1321 ggtatcattg ccgcacttgg accagatggt aagccttccc gtatcgtagt tatctacacg
     1381 acgggtagtc aggcaactat ggacgaacga aatagacaga ttgctgaaat aggggcttca
     1441 ctgattaagc attggtaaac cgatacaatt aaaggctcct tttggagcct ttttttttgg
     1501 acggaccggt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2161 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2221 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2281 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     2341 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2401 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     2461 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     2521 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     2581 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     2641 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     2701 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     2761 tttctgttca tggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     2821 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     2881 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgct
//
