LOCUS       sequence-318486-        2939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-term 6xHis 3C cleavable Amp resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2939)
  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 2939)
  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: 2 pCOLI_A_His
FEATURES             Location/Qualifiers
     source          1..2939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..97
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     331..348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    363..502
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(428..447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     588..610
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(604..795)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1032..1051
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1032..1050
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1088..1110
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1130..1147
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1160..1183
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1376..1394)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1390..1437
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1502..1593
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2458..2497
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      join(2528..2939,1..177)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
       61 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      301 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      361 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
      421 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      481 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      541 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      601 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
      661 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
      721 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
      781 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg ggtaatgata
      841 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
      901 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
      961 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttggcac catgcatcga
     1021 tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct ccggctccgg
     1081 aaataatttt gtttaacttt aagaaggaga tatacatatg ggcagcagcc atcatcatca
     1141 tcatcacagc agcggatctc tggaagttct gttccagggg cccggatccc ggtccgacat
     1201 atgagcgcgc ggaattcaaa ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga
     1261 atctagagcc tgcagtctcg acaagcttgt cgagaagtac tagaggatgt acaacgcgtg
     1321 cgcgctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1381 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1441 aaaggaggaa ctatatcggg attagcccgc ctaatgagcg ggcttttttt taattcccct
     1501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctcgtg
     1681 aaagtaaaag acgcagagga ccaattgggg gcacgagtgg gatacataga actggacttg
     1741 aatagcggta aaatccttga gagttttcgc cctgaagagc gttttccaat gatgagcact
     1801 ttcaaagttc tgctatgtgg agcagtatta tcccgtgtag atgcggggca agagcaactc
     1861 ggacgacgaa tacactattc gcagaatgac ttggttgaat actccccagt gacagaaaag
     1921 caccttacgg acggaatgac ggtaagagaa ttatgtagtg ccgccataac gatgagtgat
     1981 aacactgcgg cgaacttact tctgacaacc atcggtggac cgaaggaatt aaccgctttt
     2041 ttgcacaata tgggagacca tgtaactcgc cttgaccgtt gggaaccaga actgaatgaa
     2101 gccataccaa acgacgagcg agacaccaca atgcctgcgg caatggcaac aacattacgc
     2161 aaactattaa ctggcgaact acttactctg gcttcacggc aacaattaat agactggctt
     2221 gaagcggata aagttgcagg accactactg cgttcggcac ttcctgctgg ctggtttatt
     2281 gctgataaat ctggggcagg agagcgtggt tcacggggta tcattgccgc acttggacca
     2341 gatggtaagc cttcccgtat cgtagttatc tacacgacgg gtagtcaggc aactatggac
     2401 gaacgaaata gacagattgc tgaaataggg gcttcactga ttaagcattg gtaaaccgat
     2461 acaattaaag gctccttttg gagccttttt ttttggacgg accggtagaa aagatcaaag
     2521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2641 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgctt
//
