LOCUS       sequence-342314-        4287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial genomic integration vector, VanR repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4287)
  AUTHORS   Hao N, Chen Q, Dodd IB, Shearwin KE
  TITLE     The pIT5 Plasmid Series, an Improved Toolkit for Repeated Genome 
            Integration in E. coli.
  JOURNAL   ACS Synth Biol. 2021 Jun 30. doi: 10.1021/acssynbio.1c00215.
  PUBMED    34190535
REFERENCE   2  (bases 1 to 4287)
  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: "ACS Synth 
            Biol. 2021 Jun 30. doi: 10.1021/acssynbio.1c00215."
COMMENT     Sequence Label: pIT5_KH_VanR
FEATURES             Location/Qualifiers
     source          1..4287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        79..128
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      1121..1148
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1197..1228)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    1304..1337
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1501..1520)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1512..2327
                     /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"
     primer_bind     complement(1585..1604)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2449..2837
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2852..2885
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2915..3141
                     /gene="attP"
                     /label=HK022 attP
                     /bound_moiety="HK022 integrase"
                     /note="attachment site of phage HK022"
     terminator      complement(3176..3422)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     primer_bind     complement(3513..3535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3686..3704
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3745..3831
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3928..4014
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4111..4197
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaattccgct taacgatcgt tggctgattg gatccaattg acagctagct cagtcctagg
       61 taccattgga tccaatagct gtcaccggat gtgctttccg gtctgatgag tccgtgagga
      121 cgaaacagcc tctacaaata attttgttta atctagactc gagactagtc tcggtaccaa
      181 attccagaaa agaggcctcc cgaaaggggg gccttttttc gttttggtcc aatggcggcg
      241 cgccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
      301 attcaggggg gtgaataatg gacatgcctc gtattaaacc gggtcagcgt gttatgatgg
      361 cactgcgtaa aatgattgca agcggtgaaa tcaaaagtgg tgaacgtatt gcagaaattc
      421 cgaccgcagc agcactgggt gttagccgta tgccggttcg tatcgcactg cgttcactgg
      481 aacaagaagg tctggttgtt cgtctgggtg cacgtggtta tgcagcccgt ggtgttagca
      541 gcgatcagat tcgtgatgca attgaagttc gtggtgttct ggaaggtttt gcagcacgtc
      601 gtctggcaga acgtggtatg accgcagaaa cccatgcacg ttttgttgta ctgattgcag
      661 aaggtgaagc actgtttgca gccggtcgcc tgaatggtga agatctggat cgttatgccg
      721 catataatca ggcatttcat gataccctgg ttagcgcagc aggtaatggt gcagttgaaa
      781 gcgcactggc acgtaatggt tttgaaccgt ttgcagcagc cggtgcactg gccctggatc
      841 tgatggacct gtctgccgaa tatgaacatc tgctggcagc acatcgtcag catcaggcag
      901 ttctggatgc agttagctgt ggtgatgccg aaggtgcaga acgtattatg cgtgatcatg
      961 cactggcagc aattcgtaat gcaaaagttt ttgaagcagc agcaagcgca ggcgcaccgc
     1021 tgggtgcagc atggtcaatt cgtgcagatt gataaggatc ctaattggta acgaatcaat
     1081 gcatggcgcc taacctaaac tgacaggcat caaattaagc agaaggccat cctgacggat
     1141 ggcctttttg cgtttcgaac aattgaaaaa acctcgcgcc ttacctgttg agtaatagtc
     1201 aaaagcctcc ggtcggaggc ttttgacttt ctgcttactg aatttcggtg gtgccgttaa
     1261 ttaaccggtg ggccctcatg ataataatgg tttcttagac gtctaccgtt cgtataatgt
     1321 atgctatacg aagttatcct aggcgacaat gtaactacta gagctgatcc ttcaactcag
     1381 caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga tgttacattg
     1441 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1501 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcccgtccg cgcttaaact
     1561 ccaacatgga cgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1621 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1681 gcaaaggtag cgttgccaat gatgttacag atgagatggt ccgtctcaac tggctgacgg
     1741 agtttatgcc tctcccgacc atcaagcatt ttatccgtac tcctgatgat gcgtggttac
     1801 tcaccaccgc gattcctggg aaaacagcct tccaggtatt agaagaatat cctgattcag
     1861 gtgaaaatat tgttgatgcg ctggccgtgt tcctgcgccg gttacattcg attcctgttt
     1921 gtaattgtcc ttttaacagc gatcgtgtat ttcgtcttgc tcaggcgcaa tcacgcatga
     1981 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     2041 aagtctggaa agaaatgcac aagctcttgc cattctcacc ggattcagtc gtcactcatg
     2101 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2161 ttggacgggt cggaatcgca gaccgttacc aggaccttgc cattctttgg aactgcctcg
     2221 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2281 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaataa tactagcaga
     2341 aatcatcctt agcgaaagct aaggattttt tttatctgat taccgccttt gagtgagcgt
     2401 cgacctagtg cggccgcaag atccggccac gatgcgtccg gcgtagagga tctgaagatc
     2461 agcagttcaa cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca
     2521 tgtttaacgt actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag
     2581 ctctcatggc taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca
     2641 ataaaattaa tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa
     2701 aaaaagaata tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca
     2761 gggatgtaac gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga
     2821 acacttaacg gctgacatgg gaattaggaa gataacttcg tataatgtat gctatacgaa
     2881 cggtacatat gccatgggac aaaattgaaa tcgacaaatg attttatttt gactaataat
     2941 gacctactta cattaattta ctgataatta aagagatttt aaatatacaa cttattcacc
     3001 taaaggatga caaaataaca ttaatcactt aaaaatcatc gcattacact aatctgtggt
     3061 taaatgatag actacataat gcgacaaaac gcaacatatc cagtcactat gaatcaacta
     3121 cttagatagt attagtgacc tgagacagag cattagctag cttcttcgtc tgtttctact
     3181 ggtattggca caaacctgat tccaatttga gcaaggctat gtgccatctc gatactcgtt
     3241 cttaactcaa cagaagatgc tttgtgcata cagcccctcg tttattattt atctcctcag
     3301 ccagccgctg tgctttcagt ggatttcgga taacagaaag gccgggaaat acccagcctc
     3361 gctttgtaac ggagtagaga cgaaagtgat tgcgcctacc cggatattat cgtgaggatg
     3421 cgtcatcgcc attaattcac tgatcagtga taagctgtca aacatgagaa ttgatccggc
     3481 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3541 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3601 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3661 gctgcacatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     3721 attaattccc aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     3781 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     3841 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     3901 actgccagga attaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3961 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4021 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     4081 taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac
     4141 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4201 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4261 ccataaactg ccaggaattg gggatcg
//
