LOCUS       sequence-375749-        5335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible mCh-V5-NES-EGFP-ERBB2 NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5335)
  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 5335)
  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-ERBB2 NLS
FEATURES             Location/Qualifiers
     source          1..5335
                     /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(2105..2204)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2316..2335)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2394..2413)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2987..3520)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3709..3714,3715..3731,3732..3737))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4394..4982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4883..4902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5147..5233
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5325..5335,1..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 taccaaacga aggcagcaga
     2041 agatccggaa gtatacgatg cgtaggtgac aattggaaga ctaatgcgtt ttcgagatat
     2101 ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2161 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct
     2221 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2281 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2341 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2401 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2461 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2521 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2581 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2641 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtc
     2701 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2761 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2821 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2881 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2941 cgaggggaaa ttaggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     3001 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     3061 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     3121 cctcatgctt gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     3181 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     3241 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     3301 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     3361 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     3421 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     3481 gactgccctg ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc
     3541 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     3601 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     3661 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     3721 gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     3781 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     3841 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     3901 ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg
     3961 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     4021 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctactaat aggttgtatt
     4081 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     4141 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     4201 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4261 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4321 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4381 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4441 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4501 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4561 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4621 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4681 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4741 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4801 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4861 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4921 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4981 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5041 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgct agcatggatc
     5101 tcggggacgt ctaactacta agcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     5161 ggctcagtcg gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     5221 gagtaggaca aatccgccgg gagcggattt gaacgttgtg aagcaacggc ccggagggtg
     5281 gcgggcagga cgcccgccat aaactgccag gcatcaaact aagcagaagg ccatc
//
