LOCUS       sequence-6763-ex        2892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2892)
  AUTHORS   Nallamsetty S, Kapust RB, Tozser J, Cherry S, Tropea JE, Copeland 
            TD, Waugh DS
  TITLE     Efficient site-specific processing of fusion proteins by tobacco 
            vein mottling virus protease in vivo and in vitro.
  JOURNAL   Protein Expr Purif. 2004 Nov . 38(1):108-15.
  PUBMED    15477088
REFERENCE   2  (bases 1 to 2892)
  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 
            Expr Purif. 2004 Nov . 38(1):108-15."
COMMENT     Sequence Label: pRK1037
FEATURES             Location/Qualifiers
     source          1..2892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..61
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(82..100,101..106,107..123,124..129,130..155)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    82..100
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    107..125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             165..173
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             179..889
                     /codon_start=1
                     /product="tobacco vein mottling virus NIa protease 
                     (Nallamsetty et al., 2004)"
                     /label=TVMV protease
                     /translation="MSKALLKGVRDFNPISACVCLLENSSDGHSERLFGIGFGPYIIAN
                     QHLFRRNNGELTIKTMHGEFKVKNSTQLQMKPVEGRDIIVIKMAKDFPPFPQKLKFRQP
                     TIKDRVCMVSTNFQQKSVSSLVSESSHIVHKEDTSFWQHWITTKDGQCGSPLVSIIDGN
                     ILGIHSLTHTTNGSNYFVEFPEKFVATYLDAADGWCKNWKFNADKISWGSFTLVEDAPE
                     DDFMAKKTVAAIMD"
     terminator      922..1008
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1044..1061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1178..1723)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1258..1277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1837..1931)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1962..2756)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2073..2092)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2683..2702
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
       61 ccctttcgtc ttcacctcga gtccctatca gtgatagaga ttgacatccc tatcagtgat
      121 agagatactg agcacatcag caggacgcac tgaccgaatt gatttaagga ggtaacatat
      181 gtctaaagct ttgctgaagg gcgtgcgcga ttttaatccg atctctgctt gcgtatgcct
      241 gctggaaaac tcctcggatg gtcatagtga acgtctgttt ggcattggtt ttggcccgta
      301 tatcattgcc aaccagcatc tgtttcgtcg taacaatggc gaactgacca tcaaaaccat
      361 gcatggtgaa ttcaaagtca aaaactctac ccagctgcag atgaaaccgg ttgaaggccg
      421 tgacattatc gttatcaaaa tggctaaaga cttcccgccg ttcccgcaga aactgaaatt
      481 ccgtcagccg accatcaaag atcgtgtgtg catggtgtcc accaactttc agcagaaaag
      541 cgtctcgagc ctggtgtctg aatcctctca cattgtgcat aaagaagaca cttctttctg
      601 gcagcactgg atcaccacta aagatggcca gtgtggcagc ccactagttt ccatcattga
      661 tggcaacatt ctgggcatcc acagcctgac tcataccacc aacggtagca actacttcgt
      721 ggaatttccg gaaaaattcg tggcgactta tctagatgcc gcggatggtt ggtgcaaaaa
      781 ctggaaattc aacgcggata aaatcagctg gggttccttt accctggttg aagatgcgcc
      841 ggaagatgac ttcatggcca aaaaaactgt tgccgccatc atggactaat aaagatccca
      901 tggtacgcgt gctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      961 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag
     1021 acctagggga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     1081 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     1141 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     1201 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     1261 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     1321 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     1381 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     1441 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     1501 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     1561 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     1621 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     1681 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     1741 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
     1801 cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg
     1861 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
     1921 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     1981 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     2041 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     2101 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     2161 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     2221 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     2281 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     2341 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     2401 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     2461 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     2521 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     2581 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     2641 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     2701 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     2761 aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca gatccttggc
     2821 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     2881 gctggcaatt cc
//
