LOCUS       sequence-375653-        5475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-KPNA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5475)
  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 5475)
  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-KPNA1
FEATURES             Location/Qualifiers
     source          1..5475
                     /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(2245..2344)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2456..2475)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2534..2553)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3127..3660)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3849..3854,3855..3871,3872..3877))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4534..5122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5023..5042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5287..5373
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5465..5475,1..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 gtatgaccac
      601 cccaggaaaa gagaactttc gcctgaaaag ttacaagaac aaatctctga atcccgatga
      661 gatgcgcagg aggagggagg aagaaggact gcagttacga aagcagaaaa gagaagagca
      721 gttattcaag cggagaaatg ttgctacagc agaagaagaa acagaagaag aagttatgtc
      781 agatggaggc tttcatgagg ctcagattaa taacatggag atggcaccag gtggtgtcat
      841 cacttctgac atgattgaaa tgatattttc caaaagccca gagcaacagc tttcagcaac
      901 acagaaattc aggaagctgc tttcaaaaga acctaaccct cctattgatg aagttatcag
      961 cacaccagga gtagtggcca ggtttgtgga gttcctcaaa cgaaaagaga attgtacact
     1021 gcagtttgaa tcagcttggg tactgacaaa tattgcttca ggaaattctc ttcagacccg
     1081 aattgtgatt caggcaggag ctgtgcccat cttcatagag ttgctcagct cagagtttga
     1141 agatgtccag gaacaggcag tctgggctct tggcaacatt gctggagata gtaccatgtg
     1201 cagggactat gtcttagact gcaatatcct tccccctctt ttgcagttat tttcaaagca
     1261 aaaccgcctg accatgaccc ggaatgcagt atgggctttg tctaatctct gtagagggaa
     1321 aagtccacct ccagaatttg caaaggtttc tccatgtctg aatgtgcttt cctggttgct
     1381 gtttgtcagt gacactgatg tactggctga tgcctgctgg gccctctcat atctatcaga
     1441 tggacccaat gataaaattc aagcggtcat cgatgcggga gtatgtagga gacttgtgga
     1501 actgctgatg cataatgatt ataaagtggt ttctcctgct ttgcgagctg tgggaaacat
     1561 tgtcacaggg gatgatattc agacacaggt aattctgaat tgctcagctc tgcagagttt
     1621 attgcatttg ctgagtagcc caaaggaatc tatcaaaaag gaagcatgtt ggacgatatc
     1681 taatattaca gctggaaata gggcacagat ccagactgtg atagatgcca acattttccc
     1741 agccctcatt agtattttac aaactgctga atttcggaca agaaaagaag cagcttgggc
     1801 catcacaaat gcaacttctg gaggatcagc tgaacagatc aagtacctag tagaactggg
     1861 ttgtatcaag ccgctctgtg atctcctcac ggtcatggac tctaagattg tacaggttgc
     1921 cctaaatggc ttggaaaata tcctgaggct tggagaacag gaagccaaaa ggaatggcac
     1981 tggcattaac ccttactgtg ctttgattga agaagcttat ggtctggata aaattgagtt
     2041 cttacagagt catgaaaacc aggagatcta ccaaaaggcc tttgatctta ttgagcatta
     2101 cttcgggacc gaagatgaag acagcagcat tgcaccccag gttgacctta accagcagca
     2161 gtacatcttc caacagtgtg aggctcctat ggaaggtttc cagctttgac aattggaaga
     2221 ctaatgcgtt ttcgagatat ctagacccag ctttcttgta caaagttggc attataagaa
     2281 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     2341 tttgccatcc agctgcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     2401 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2461 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     2521 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     2581 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     2641 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     2701 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     2761 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     2821 gttgatgcgc tggcagtgtc cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     2881 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     2941 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     3001 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     3061 cttgataacc ttatttttga cgaggggaaa ttaggccggg aagccgatct cggcttgaac
     3121 gaattgttag gtggcggtac ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc
     3181 caactttgta tagagagcca ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac
     3241 ataagcacca agcgcgttgg cctcatgctt gaggagattg atgagcgcgg tggcaatgcc
     3301 ctgcctccgg tgctcgccgg agactgcgag atcatagata tagatctcac tacgcggctg
     3361 ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca accgcttctt ggtcgaaggc
     3421 agcaagcgcg atgaatgtct tactacggag caagttcccg aggtaatcgg agtccggctg
     3481 atgttgggag taggtggcta cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg
     3541 catggatttg acttggtcag ggccgagcct acatgtgcga atgatgccca tacttgagcc
     3601 acctaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat
     3661 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     3721 cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc
     3781 cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact
     3841 tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt
     3901 tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca tttctgtcct
     3961 ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg gcggccttgc
     4021 tgttcttcta cggcaaggtg ctgtgcacgg atctgccctg gcttcaggag atcggaagac
     4081 ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg
     4141 gttttctgga aggcgagcat cgtttgttcg cccaggactc tagctatagt tctagtggtt
     4201 ggctactaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4261 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4321 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4381 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     4441 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4561 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4621 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4681 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4741 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4801 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4861 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4921 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4981 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5041 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5101 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5161 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5221 tattaccgct agcatggatc tcggggacgt ctaactacta agcgagagta gggaactgcc
     5281 aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg cctttcgttt tatctgttgt
     5341 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgtg
     5401 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaact
     5461 aagcagaagg ccatc
//
