LOCUS       sequence-375786-        6489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-KPNB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6489)
  AUTHORS   Wang L, Paudel BB, McKnight RA, Janes KA
  TITLE     Nucleocytoplasmic transport of active HER2 causes fractional escape 
            from the DCIS-like state.
  JOURNAL   Nat Commun. 2023 Apr 13;14(1):2110. doi: 10.1038/s41467-023-37914-x.
  PUBMED    37055441
REFERENCE   2  (bases 1 to 6489)
  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: "Nat 
            Commun."; date: "2023-04-13"; volume: "14(1):2110. doi"; pages: " 
            10.1038/s41467-023-37914-x"
COMMENT     Sequence Label: pEN_TT 3xFLAG-KPNB1
FEATURES             Location/Qualifiers
     source          1..6489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..48
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    67..166
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        198..512
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    206..224
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    242..260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    277..295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    349..367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    384..402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    420..438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     485..509
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             521..586
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(3259..3358)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3470..3489)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3548..3567)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4141..4674)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4863..4868,4869..4885,4886..4891))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5548..6136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6037..6056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      6301..6387
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(6479..6489,1..17)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc
       61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat
      121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat
      181 tcagtcgact ggatcccgag tttactccct atcagtgata gagaacgtat gtcgagttta
      241 ctccctatca gtgatagaga acgatgtcga gtttactccc tatcagtgat agagaacgta
      301 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      361 atagagaacg tatgtcgagt ttatccctat cagtgataga gaacgtatgt cgagtttact
      421 ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt acggtgggag gcctatataa
      481 gcagagctcg tttagtgaac cgtcagatcg ccactagatg gactacaaag accatgacgg
      541 tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagacta gaatggagct
      601 gatcaccatt ctcgagaaga ccgtgtctcc cgatcggctg gagctggaag cggcgcagaa
      661 gttcctggag cgtgcggccg tggagaacct gcccactttc cttgtggaac tgtccagagt
      721 gctggcaaat ccaggaaaca gtcaggttgc cagagttgca gctggtctac aaatcaagaa
      781 ctctttgaca tctaaagatc cagatatcaa ggcacaatat cagcagaggt ggcttgctat
      841 tgatgctaat gctcgacgag aagtcaagaa ctatgttttg cagacattgg gtacagaaac
      901 ttaccggcct agttctgcct cacagtgtgt ggctggtatt gcttgtgcag agatcccagt
      961 aaaccagtgg ccagaactca ttcctcagct ggtggccaat gtcacaaacc ccaacagcac
     1021 agagcacatg aaggagtcga cattggaagc catcggttat atttgccaag atatagaccc
     1081 agagcagcta caagataaat ccaatgagat tctgactgcc ataatccagg ggatgaggaa
     1141 agaagagcct agtaataatg tgaagctagc tgctacgaat gcactcctga actcattgga
     1201 gttcaccaaa gcaaactttg ataaagagtc tgaaaggcac tttattatgc aggtggtctg
     1261 tgaagccaca cagtgtccag atacgagggt acgagtggct gctttacaga atctggtgaa
     1321 gataatgtcc ttatattatc agtacatgga gacatatatg ggtcctgctc tttttgcaat
     1381 cacaatcgaa gcaatgaaaa gtgacattga tgaggtggct ttacaaggga tagaattctg
     1441 gtccaatgtc tgtgatgagg aaatggattt ggccattgaa gcttcagagg cagcagaaca
     1501 aggacggccc cctgagcaca ccagcaagtt ttatgcgaag ggagcactac agtatctggt
     1561 tccaatcctc acacagacac taactaaaca ggacgaaaat gatgatgacg atgactggaa
     1621 cccctgcaaa gcagcagggg tgtgcctcat gcttctggcc acctgctgtg aagatgacat
     1681 tgtcccacat gtcctcccct tcattaaaga acacatcaag aacccagatt ggcggtaccg
     1741 ggatgcagca gtgatggctt ttggttgtat cttggaagga ccagagccca gtcagctcaa
     1801 accactagtt atacaggcta tgcccaccct aatagaatta atgaaagacc ccagtgtagt
     1861 tgttcgagat acagctgcat ggactgtagg cagaatttgt gagctgcttc ctgaagctgc
     1921 catcaatgat gtctacttgg ctcccctgct acagtgtctg attgagggtc tcagtgctga
     1981 acccagagtg gcttcaaatg tgtgctgggc tttctccagt ctggctgaag ctgcttatga
     2041 agctgcagac gttgctgatg atcaggaaga accagctact tactgcttat cttcttcatt
     2101 tgaactcata gttcagaagc tcctagagac tacagacaga cctgatggac accagaacaa
     2161 cctgagaagt tctgcatatg aatctctgat ggaaattgtg aaaaacagtg ccaaggattg
     2221 ttatcctgct gtccagaaaa cgactttggt catcatggaa cgactgcaac aggttcttca
     2281 gatggagtca catatccaga gcacatccga tagaatccag ttcaatgacc ttcagtcttt
     2341 actctgtgca actcttcaga atgttcttcg gaaagtgcaa catcaagatg ctttgcagat
     2401 ctctgatgtg gttatggcct ccctgttaag gatgttccaa agcacagctg ggtctggggg
     2461 agtacaagag gatgccctga tggcagttag cacactggtg gaagtgttgg gtggtgaatt
     2521 cctcaagtac atggaggcct ttaaaccctt cctgggcatt ggattaaaaa attatgctga
     2581 ataccaggtt tgtttggcag ctgtgggctt agtgggagac ttgtgccgtg ccctgcaatc
     2641 caacatcata cctttctgtg acgaggtgat gcagctgctt ctggaaaatt tggggaatga
     2701 gaacgtccac aggtctgtga agccgcagat tctgtcagtg tttggtgata ttgcccttgc
     2761 tattggagga gagtttaaaa aatacttaga ggttgtattg aatactcttc agcaggcctc
     2821 ccaagcccag gtggacaagt cagactatga catggtggat tatctgaatg agctaaggga
     2881 aagctgcttg gaagcctata ctggaatcgt ccagggatta aagggggatc aggagaacgt
     2941 acacccggat gtgatgctgg tacaacccag agtagaattt attctgtctt tcattgacca
     3001 cattgctgga gatgaggatc acacagatgg agtagtagct tgtgctgctg gactaatagg
     3061 ggacttatgt acagcatttg ggaaggatgt actgaaatta gtagaagcta ggccaatgat
     3121 ccatgaattg ttaactgaag ggcggagatc gaagactaac aaagcaaaaa cccttgctac
     3181 atgggcaaca aaagaactga ggaaactgaa gaaccaagct tgacaattgg aagactaatg
     3241 cgttttcgag atatctagac ccagctttct tgtacaaagt tggcattata agaaagcatt
     3301 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     3361 atccagctgc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     3421 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     3481 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     3541 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     3601 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     3661 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     3721 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     3781 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     3841 gcgctggcag tgtccctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     3901 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     3961 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4021 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4081 aaccttattt ttgacgaggg gaaattaggc cgggaagccg atctcggctt gaacgaattg
     4141 ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt
     4201 tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc
     4261 accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct
     4321 ccggtgctcg ccggagactg cgagatcata gatatagatc tcactacgcg gctgctcaaa
     4381 cttgggcaga acgtaagccg cgagagcgcc aacaaccgct tcttggtcga aggcagcaag
     4441 cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg
     4501 ggagtaggtg gctacgtctc cgaactcacg accgaaaaga tcaagagcag cccgcatgga
     4561 tttgacttgg tcagggccga gcctacatgt gcgaatgatg cccatacttg agccacctaa
     4621 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctgcgta acatcgttgc
     4681 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
     4741 gcatagactg tacaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac
     4801 caccgctgcg ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt
     4861 acagtttacg aaccgaacag gcttatgtca actgggttcg tgccttcatc cgtttccacg
     4921 gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg
     4981 cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct
     5041 tctacggcaa ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc
     5101 cgtcgcggcg cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc
     5161 tggaaggcga gcatcgtttg ttcgcccagg actctagcta tagttctagt ggttggctac
     5221 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
     5281 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
     5341 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
     5401 tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg acttgacggg
     5461 acggcgcaag ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6241 cgctagcatg gatctcgggg acgtctaact actaagcgag agtagggaac tgccaggcat
     6301 caaataaaac gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg
     6361 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa
     6421 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag
     6481 aaggccatc
//
