LOCUS       sequence-310332-        2608 bp    DNA     circular SYN 15-APR-2025
DEFINITION  N-term HA 3C cleavable Kan resistant bacterial expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2608)
  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 2608)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: 35 pCOLI_K_HA
FEATURES             Location/Qualifiers
     source          1..2608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     317..336
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(410..501)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(517..1266)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(527..544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(767..786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1462..1481
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1462..1480
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1518..1540
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1560..1577
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1590..1616
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1623..1649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1656..1682
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1695..1718
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1911..1929)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1925..1972
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2202..2608,1..409))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(join(2202..2608,1..409))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
       61 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
      121 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
      181 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
      241 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
      301 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
      361 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata ctcttccttt
      421 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      481 gtatttagaa aaataaacaa ataggcatgc agcgctcttc cgcttcctcg ctcactgact
      541 cgctacgctc ggtcgttcga ctgcggcgag cggtgtcagc tcactcaaaa gcggtaatac
      601 ggttatccac agaatcaggg gataaagccg gaaagaacat gtgagcaaaa agcaaagcac
      661 cggaagaagc caacgccgca ggcgtttttc cataggctcc gcccccctga cgagcatcac
      721 aaaaatcgac gctcaagcca gaggtggcga aacccgacag gactataaag ataccaggcg
      781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgttggtat
      901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1021 ttatcgccac tggcagcagc cattggtaac tgatttagag gactttgtct tgaagttatg
     1081 cacctgttaa ggctaaactg aaagaacaga ttttggtgag tgcggtcctc caacccactt
     1141 accttggttc aaagagttgg tagctcagcg aaccttgaga aaaccaccgt tggtagcggt
     1201 ggtttttctt tatttatgag atgatgaatc aatcggtcta tcaagtcaac gaacagctat
     1261 tccgttactc tagatttcag tgcaatttat ctcttcaaat gtagcacctg aagtcagccc
     1321 catacgatat aagttgtaat tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg
     1381 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     1441 cttacattaa ttgcgttgcg ctaatacgac tcactatagg gagaccacaa cggtttccct
     1501 ccggctccgg aaataatttt gtttaacttt aagaaggaga tatacatatg gggagttcgc
     1561 accatcatca ccaccatagc tctggctcct atccgtatga cgtgccagac tatgctggta
     1621 gctatcctta cgatgtcccc gattatgcgg gatcttatcc atacgatgtg ccagactatg
     1681 caagttcagg ttccctggaa gttctgttcc aggggcccgg atcccggtcc gacatatgag
     1741 cgcgcggaat tcaaaggcct acgtcgacga gctcactagt cgcggccgct ttcgaatcta
     1801 gagcctgcag tctcgacaag cttgtcgaga agtactagag gatgtacaac gcgtgcgcgc
     1861 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1921 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     1981 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     2041 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc
     2101 gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2161 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     2221 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2281 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2341 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2401 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2461 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2521 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2581 acgcggtcgc tgttaaaagg acaattac
//
