LOCUS       sequence-342311-        4456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial genomic integration vector, NahR repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4456)
  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 4456)
  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_KD_NahR
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        145..194
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     terminator      1354..1381
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1430..1461)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    1537..1570
                     /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(1734..1753)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1745..2560
                     /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(1818..1837)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2682..3070
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    3085..3118
                     /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)."
     terminator      complement(3345..3591)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     primer_bind     complement(3682..3704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3855..3873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3914..4000
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4097..4183
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4280..4366
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gaattccgct taacgatcgt tggctggggg cctcgcttgg gttattgctg gtgcccggcc
       61 gggcgcaata ttcatgttga tgatttatta tatatcgagt ggtgtattta tttatattgt
      121 ttgctccgtt accgttatta acagctgtca ccggatgtgc tttccggtct gatgagtccg
      181 tgaggacgaa acagcctcta caaataattt tgtttaatct agactcgaga ctagtctcgg
      241 taccaaattc cagaaaagag gcctcccgaa aggggggcct ttttcgtttt ggtccaatgg
      301 cggcgcgcca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag
      361 agtcaattca tgggggttga atatggaact gcgtgacctg gatttaaacc tgctggtggt
      421 gttcaaccag ttgctggtcg acagacgcgt ctctgtcact gcggagaacc tgggcctgac
      481 ccagcctgcc gtgagcaatg cgctgaaacg cctgcgcacc tcgctacagg acccactctt
      541 cgtgcgcaca catcagggaa tggaacccac accctatgcc gcgcatctgg ccgagcacgt
      601 cacttcggcc atgcacgcac tgcgcaacgc cctacagcac catgaaagct tcgatccgct
      661 gaccagcgag cgtaccttca ccctggccat gaccgacatt ggcgagatct acttcatgcc
      721 gcggctgatg gatgcgctgg ctcaccaggc ccccaattgc gtgatcagta cggtgcgcga
      781 cagttcgatg agcctgatgc aggccttgca gaacggaacc gtggacttgg ccgtgggcct
      841 gcttcccaat ctgcaaactg gcttctttca gcgccggctg ctccagaatc actacgtgtg
      901 cctatgtcgc aaggaccatc cagtcacccg cgaacccctg actctggagc gcttctgttc
      961 ctacggccac gtgcgtgtca tcgccgctgg caccggccac ggcgaggtgg acacgtacat
     1021 gacacgggtc ggcatccggc gcgacatccg tctggaagtg ccgcacttcg ccgccgttgg
     1081 ccacatcctc cagcgcaccg atctgctcgc cactgtgccg atatgtttag ccgactgctg
     1141 cgtagagccc ttcggcctaa gcgccttgcc gcacccagtc gtcttgcctg aaatagccat
     1201 caacatgttc tggcatgcga agtaccacaa ggacctagcc aatatttggt tgcggcaact
     1261 gatgtttgac ctgtttacgg attgataagg atcctaattg gtaacgaatc aatgcatggc
     1321 gcctaaccta aactgacagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1381 ttgcgtttcg aacaattgaa aaaacctcgc gccttacctg ttgagtaata gtcaaaagcc
     1441 tccggtcgga ggcttttgac tttctgctta ctgaatttcg gtggtgccgt taattaaccg
     1501 gtgggccctc atgataataa tggtttctta gacgtctacc gttcgtataa tgtatgctat
     1561 acgaagttat cctaggcgac aatgtaacta ctagagctga tccttcaact cagcaaaagt
     1621 tcgatttatt caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     1681 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     1741 tgttatgagc catattcaac gggaaacgtc ttgctcccgt ccgcgcttaa actccaacat
     1801 ggacgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1861 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1921 tagcgttgcc aatgatgtta cagatgagat ggtccgtctc aactggctga cggagtttat
     1981 gcctctcccg accatcaagc attttatccg tactcctgat gatgcgtggt tactcaccac
     2041 cgcgattcct gggaaaacag ccttccaggt attagaagaa tatcctgatt caggtgaaaa
     2101 tattgttgat gcgctggccg tgttcctgcg ccggttacat tcgattcctg tttgtaattg
     2161 tccttttaac agcgatcgtg tatttcgtct tgctcaggcg caatcacgca tgaataacgg
     2221 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2281 gaaagaaatg cacaagctct tgccattctc accggattca gtcgtcactc atggtgattt
     2341 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2401 ggtcggaatc gcagaccgtt accaggacct tgccattctt tggaactgcc tcggtgagtt
     2461 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2521 taaattgcag tttcatttga tgctcgatga gtttttctaa taatactagc agaaatcatc
     2581 cttagcgaaa gctaaggatt ttttttatct gattaccgcc tttgagtgag cgtcgaccta
     2641 gtgcggccgc aagatccggc cacgatgcgt ccggcgtaga ggatctgaag atcagcagtt
     2701 caacctgttg atagtacgta ctaagctctc atgtttcacg tactaagctc tcatgtttaa
     2761 cgtactaagc tctcatgttt aacgaactaa accctcatgg ctaacgtact aagctctcat
     2821 ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga acaataaaat
     2881 taatataaat cagcaactta aatagcctct aaggttttaa gttttataag aaaaaaaaga
     2941 atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag ccagggatgt
     3001 aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca ggaacactta
     3061 acggctgaca tgggaattag gaagataact tcgtataatg tatgctatac gaacggtaca
     3121 tatgccatgg tgtggtcaat gtgtggacgt gaccagaaac aaatcctttt atttcatgta
     3181 attagatatt aaaaaatgtc accaataact gacttggtag cccccgccgt agcggtattt
     3241 tctgttgtat tacaattagt taaattactc atatcgcttc aattggctct agttaactct
     3301 ggtttatgga cgcattatgg acatgctagc ttcttcgtct gtttctactg gtattggcac
     3361 aaacctgatt ccaatttgag caaggctatg tgccatctcg atactcgttc ttaactcaac
     3421 agaagatgct ttgtgcatac agcccctcgt ttattattta tctcctcagc cagccgctgt
     3481 gctttcagtg gatttcggat aacagaaagg ccgggaaata cccagcctcg ctttgtaacg
     3541 gagtagagac gaaagtgatt gcgcctaccc ggatattatc gtgaggatgc gtcatcgcca
     3601 ttaattcact gatcagtgat aagctgtcaa acatgagaat tgatccggct gcctcgcgcg
     3661 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3721 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3781 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtatg ctgcacatga
     3841 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttaattccca
     3901 attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3961 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     4021 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa
     4081 ttaattcccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4141 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     4201 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     4261 gaattaattc cccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4321 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     4381 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     4441 caggaattgg ggatcg
//
