LOCUS       sequence-375753-        5314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible mCh-V5-NES-EGFP-SV40 NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5314)
  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 5314)
  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 mCh-V5-NES-EGFP-SV40 NLS
FEATURES             Location/Qualifiers
     source          1..5314
                     /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             519..1226
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(664..682)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     932..951
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1227..1268
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(1305..1307,1308..1310,1311..2018)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(1350..1371)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1611..1630)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1958..1979
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2025..2045
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(2084..2183)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2295..2314)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2373..2392)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2966..3499)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3688..3693,3694..3710,3711..3716))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4373..4961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4862..4881
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5126..5212
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5304..5314,1..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 ccactagtat ggtgagcaag ggcgaggagg
      541 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
      601 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
      661 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
      721 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
      781 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
      841 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
      901 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
      961 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     1021 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     1081 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     1141 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     1201 ccggcggcat ggacgagctg tacaagggta agcctatccc taaccctctc ctcggtctcg
     1261 attctacgac cggtttagcc ttgaaattag caggtcttga tatcatggtg agcaagggcg
     1321 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1381 acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1441 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1501 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     1561 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1621 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1681 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1741 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggcgaact
     1801 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1861 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1921 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1981 ccgccgccgg gatcactctc ggcatggacg agctgtacgg taccccaaaa aagaagagaa
     2041 aggtatgaca attggaagac taatgcgttt tcgagatatc tagacccagc tttcttgtac
     2101 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     2161 agtcaaaata aaatcattat ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct
     2221 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2281 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2341 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2401 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2461 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2521 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2581 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2641 tgattcaggt gaaaatattg ttgatgcgct ggcagtgtcc ctgcgccggt tgcattcgat
     2701 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2761 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2821 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2881 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taggccggga
     2941 agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca
     3001 tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat
     3061 ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga
     3121 tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat
     3181 agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa
     3241 ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga
     3301 ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga
     3361 aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa
     3421 tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat
     3481 cgttgctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     3541 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
     3601 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     3661 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg
     3721 gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa
     3781 gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt
     3841 caggcattgg cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg
     3901 cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat
     3961 gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct
     4021 agctatagtt ctagtggttg gctactaata ggttgtattg atgttggacg agtcggaatc
     4081 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4141 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4201 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     4261 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     4321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4381 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4441 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4501 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4561 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4621 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4681 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4741 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4801 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4861 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4921 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4981 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5041 ctgattctgt ggataaccgt attaccgcta gcatggatct cggggacgtc taactactaa
     5101 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc
     5161 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     5221 agcggatttg aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     5281 aactgccagg catcaaacta agcagaaggc catc
//
