LOCUS       sequence-375709-        5503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible mCh-V5-NES-EGFP-IBB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5503)
  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 5503)
  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-IBB
FEATURES             Location/Qualifiers
     source          1..5503
                     /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"
     protein_bind    complement(2273..2372)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2484..2503)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2562..2581)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3155..3688)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3877..3882,3883..3899,3900..3905))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4562..5150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5051..5070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5315..5401
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5493..5503,1..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 taccatgtcc accaacgaga
     2041 atgctaatac accagctgcc cgtcttcaca gattcaagaa caagggaaaa gacagtacag
     2101 aaatgaggcg tcgcagaata gaggtcaatg tggagctgag gaaagctaag aaggatgacc
     2161 agatgctgaa gaggagaaat gtaagctcat ttcctgatga tgctacttct ccgctgcagg
     2221 aaaaccgcaa caactaacaa ttggaagact aatgcgtttt cgagatatct agacccagct
     2281 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     2341 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgcagctct ggcccgtgtc
     2401 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2461 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2521 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2581 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2641 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2701 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2761 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2821 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgtccc tgcgccggtt
     2881 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2941 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3001 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3061 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3121 aggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     3181 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     3241 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     3301 ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     3361 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     3421 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     3481 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     3541 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     3601 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     3661 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     3721 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     3781 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     3841 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     3901 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     3961 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
     4021 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat
     4081 ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg
     4141 accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc
     4201 caggactcta gctatagttc tagtggttgg ctactaatag gttgtattga tgttggacga
     4261 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     4321 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     4381 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     4441 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     4501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4681 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5221 cgttatcccc tgattctgtg gataaccgta ttaccgctag catggatctc ggggacgtct
     5281 aactactaag cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgga
     5341 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     5401 tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc gggcaggacg
     5461 cccgccataa actgccaggc atcaaactaa gcagaaggcc atc
//
