LOCUS       sequence-375752-        5454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-KPNA2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5454)
  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 5454)
  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-KPNA2
FEATURES             Location/Qualifiers
     source          1..5454
                     /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(2224..2323)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2435..2454)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2513..2532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3106..3639)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3828..3833,3834..3850,3851..3856))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4513..5101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5002..5021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5266..5352
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5444..5454,1..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 gtatgtccac
      601 caacgagaat gctaatacac cagctgcccg tcttcacaga ttcaagaaca agggaaaaga
      661 cagtacagaa atgaggcgtc gcagaataga ggtcaatgtg gagctgagga aagctaagaa
      721 ggatgaccag atgctgaaga ggagaaatgt aagctcattt cctgatgatg ctacttctcc
      781 gctgcaggaa aaccgcaaca accagggcac tgtaaattgg tctgttgatg acattgtcaa
      841 aggcataaat agcagcaatg tggaaaatca gctccaagct actcaagctg ccaggaaact
      901 actttccaga gaaaaacagc cccccataga caacataatc cgggctggtt tgattccgaa
      961 atttgtgtcc ttcttgggca gaactgattg tagtcccatt cagtttgaat ctgcttgggc
     1021 actcactaac attgcttctg ggacatcaga acaaaccaag gctgtggtag atggaggtgc
     1081 catcccagca ttcatttctc tgttggcatc tccccatgct cacatcagtg aacaagctgt
     1141 ctgggctcta ggaaacattg caggtgatgg ctcagtgttc cgagacttgg ttattaagta
     1201 cggtgcagtt gacccactgt tggctctcct tgcagttcct gagatgtcat ctttagcatg
     1261 tggctactta cgtaatctta cctggacact ttctaatctt tgccgcaaca agaatcctgc
     1321 acccccgata gatgctgttg agcagattct tcctacctta gttcggctcc tgcatcatga
     1381 tgatccagaa gtgttagcag atacctgctg ggctatttcc taccttactg atggtccaaa
     1441 tgaacgaatt ggcatggtgg tgaaaacagg agttgtgccc caacttgtga agcttctagg
     1501 agcttctgaa ttgccaattg tgactcctgc cctaagagcc atagggaata ttgtcactgg
     1561 tacagatgaa cagactcagg ttgtgattga tgcaggagca ctcgccgtct ttcccagcct
     1621 gctcaccaac cccaaaacta acattcagaa ggaagctacg tggacaatgt caaacatcac
     1681 agccggccgc caggaccaga tacagcaagt tgtgaatcat ggattagtcc cattccttgt
     1741 cagtgttctc tctaaggcag attttaagac acaaaaggaa gctgtgtggg ccgtgaccaa
     1801 ctataccagt ggtggaacag ttgaacagat tgtgtacctt gttcactgtg gcataataga
     1861 accgttgatg aacctcttaa ctgcaaaaga taccaagatt attctggtta tcctggatgc
     1921 catttcaaat atctttcagg ctgctgagaa actaggtgaa actgagaaac ttagtataat
     1981 gattgaagaa tgtggaggct tagacaaaat tgaagctcta caaaaccatg aaaatgagtc
     2041 tgtgtataag gcttcgttaa gcttaattga gaagtatttc tctgtagagg aagaggaaga
     2101 tcaaaacgtt gtaccagaaa ctacctctga aggctacact ttccaagttc aggatggggc
     2161 tcctgggacc tttaactttt gagcatgcca attggaagac taatgcgttt tcgagatatc
     2221 tagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2281 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagctc
     2341 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     2401 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2461 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2521 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     2581 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2641 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2701 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     2761 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgtcc
     2821 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2881 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2941 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     3001 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3061 gaggggaaat taggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact
     3121 tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac
     3181 tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc
     3241 ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga
     3301 gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta
     3361 agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt
     3421 actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac
     3481 gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg
     3541 gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg
     3601 actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc cataacatca
     3661 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtacaa
     3721 aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg
     3781 tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt ttacgaaccg
     3841 aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg
     3901 caaccttggg cagcagcgaa gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg
     3961 tttcggtctc cacgcatcgt caggcattgg cggccttgct gttcttctac ggcaaggtgc
     4021 tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc
     4081 cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc
     4141 gtttgttcgc ccaggactct agctatagtt ctagtggttg gctactaata ggttgtattg
     4201 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     4261 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     4321 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     4381 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     4441 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4501 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4561 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4621 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4681 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4741 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4801 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4861 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4921 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4981 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5041 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5101 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5161 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcta gcatggatct
     5221 cggggacgtc taactactaa gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     5281 gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     5341 agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg
     5401 cgggcaggac gcccgccata aactgccagg catcaaacta agcagaaggc catc
//
