LOCUS       sequence-319357-        3227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 6xHis SUMO 3C cleavable Amp resistant bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3227)
  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 3227)
  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: 19 pCOLI_A_SUMO
FEATURES             Location/Qualifiers
     source          1..3227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      29..617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     518..537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     771..788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    803..942
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(868..887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1028..1050
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1044..1235)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1472..1491
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1472..1490
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1528..1550
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1570..1587
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1588..1875
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             1888..1911
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2104..2122)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2118..2165
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2230..2321
                     /gene="bla"
                     /label=AmpR promoter
     terminator      3186..3225
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
ORIGIN
        1 gaccggtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
       61 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
      181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      661 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      721 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
      781 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
      841 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      901 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
      961 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     1021 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     1081 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     1141 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     1201 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     1261 ctgttcatgg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     1321 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     1381 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     1441 gtgttggcac catgcatcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
     1501 cggtttccct ccggctccgg aaataatttt gtttaacttt aagaaggaga tatacatatg
     1561 ggcagcagcc atcatcatca tcatcactcg gactcagaag tcaatcaaga agctaagcca
     1621 gaggtcaagc cagaagtcaa gcctgagact cacatcaatt taaaggtgtc cgatggatct
     1681 tcagagatct tcttcaagat caaaaagacc actcctttaa gaaggctgat ggaagcgttc
     1741 gctaaaagac agggtaagga aatggactcc ttaagattct tgtacgacgg tattagaatt
     1801 caagctgatc agacccctga agatttggac atggaggata acgatattat tgaggctcac
     1861 agagaacaga ttggtagcag cggatctctg gaagttctgt tccaggggcc cggatcccgg
     1921 tccgacatat gagcgcgcgg aattcaaagg cctacgtcga cgagctcact agtcgcggcc
     1981 gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatgtac
     2041 aacgcgtgcg cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     2101 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2161 ttttgctgaa aggaggaact atatcgggat tagcccgcct aatgagcggg ctttttttta
     2221 attcccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2281 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2401 cgctcgtgaa agtaaaagac gcagaggacc aattgggggc acgagtggga tacatagaac
     2461 tggacttgaa tagcggtaaa atccttgaga gttttcgccc tgaagagcgt tttccaatga
     2521 tgagcacttt caaagttctg ctatgtggag cagtattatc ccgtgtagat gcggggcaag
     2581 agcaactcgg acgacgaata cactattcgc agaatgactt ggttgaatac tccccagtga
     2641 cagaaaagca ccttacggac ggaatgacgg taagagaatt atgtagtgcc gccataacga
     2701 tgagtgataa cactgcggcg aacttacttc tgacaaccat cggtggaccg aaggaattaa
     2761 ccgctttttt gcacaatatg ggagaccatg taactcgcct tgaccgttgg gaaccagaac
     2821 tgaatgaagc cataccaaac gacgagcgag acaccacaat gcctgcggca atggcaacaa
     2881 cattacgcaa actattaact ggcgaactac ttactctggc ttcacggcaa caattaatag
     2941 actggcttga agcggataaa gttgcaggac cactactgcg ttcggcactt cctgctggct
     3001 ggtttattgc tgataaatct ggggcaggag agcgtggttc acggggtatc attgccgcac
     3061 ttggaccaga tggtaagcct tcccgtatcg tagttatcta cacgacgggt agtcaggcaa
     3121 ctatggacga acgaaataga cagattgctg aaataggggc ttcactgatt aagcattggt
     3181 aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacg
//
