LOCUS       sequence-375758-        7425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible NPIPB11-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7425)
  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 7425)
  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 NPIPB11-FLAG
FEATURES             Location/Qualifiers
     source          1..7425
                     /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"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3909..3932
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3952..3969)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3958..4069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4195..4294)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4406..4425)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4484..4503)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(5077..5610)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5799..5804,5805..5821,5822..5827))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      6484..7072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6973..6992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      7237..7323
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(7415..7425,1..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 ccactagtgg taccgagctc ggatccgcca
      541 ccatggtgaa gctctctatt gtcctgaccc cacagttcct gtcccatgac cagggccagc
      601 tcaccaagga gctgcagcag cacgtaaagt cagtgacatg cccatgcgag tacctgagga
      661 aggttatcaa tactctggct gaccatcatc atcgtgggac tgactttggt ggaagtcctt
      721 ggttacatat tattattgcg tttccgacaa gttataaagt tgtcattacc ctctggatag
      781 tttacctttg ggtgtctctc ctgaagacta tcttctggtc tcgaaatgga catgatggat
      841 ccacggatgt acagcagaga gcctggaggt ccaaccgccg tagacaggaa gggctgaggt
      901 ccatttgtat gcacacaaag aaaagagttt cttcctttcg aggaaataaa attggcctga
      961 aagacgtcat tactctacgg aggcatgtgg aaacaaaagt tagagctaaa atccgtaaga
     1021 ggaaggtgac aacgaaaatc aaccgtcatg acaaaatcaa tggaaagagg aagaccgcca
     1081 gaaaacagaa aatgtttcaa cgtgcgcaag agttgcggcg gcgggcagag gactaccaca
     1141 aatgcaaaat ccccccttct gcaagaaagg ctctttgcaa ctgggtcaga atggcggcag
     1201 cggagcatcg tcattcttca ggattgccct actggcccta cctcacagct gaaactttaa
     1261 aaaacaggat gggccaccag ccacctcctc caactcaaca acattgtata actgataact
     1321 ccctgagcct caagacacct ctcgagtgtc tgctcactcc ccttccaccc tcagcggatg
     1381 ataatctcaa gacacctccc gagtgtctgc tcactcccct tccaccctca gcggatgata
     1441 atctcaagac acctcccgag tgtctgctca ctccccttcc accctcagct ccaccctcag
     1501 ctccaccctc agcggatgat aatctcaaga cacgtgccga gtgtctgctc catccccttc
     1561 caccctcagc ggatgataat ctcaagacac cttccgagcg tcagctcact ccccttccac
     1621 cctcagctcc accctcagca gatgataata tcaagacaac tgccgagcgt ctgcgggggc
     1681 cgcttccacc ctcagcggat gataatctca agacaccttc tgagcgtcag ctcactcccc
     1741 ttccaccctc agctccaccc tcagcagatg ataatatcaa gacacctgcc gagcatctgc
     1801 gggggccgct tccaccctca gcggatgata atctcaagac accttctgag cgtcagctca
     1861 ctccccttcc accctcagct ccaccctcag cagatgataa tatcaagaca cctgccgagc
     1921 gtctgcgggg gccgcttcca ccctcagccg atgataatct caagacacct tctgagcgtc
     1981 agctcactcc ccttccaccc tcagctccac cctcagcaga tgataatatc aagacacctg
     2041 ccgagcatct gcgggggccg cttccaccct cagcggatga taatctcaag acaccttctg
     2101 agcgtcagct cactcccctt ccaccctcag ctccaccctc agcagatgat aatatcaaga
     2161 caactgccga gcatctgcgg gggccgcttc caccctcagc ggatgataat ctcaagacac
     2221 cttccgagcg tcagctcact ccccttccac cctcagctcc accctcagca gatgataata
     2281 tcaagacacc tgccgagcat ctgcagttcc gcttccaccc tcagcggatg ataatctcaa
     2341 gagaccttcc gagcgtcagc tcactcccct tccaccctca gctccaccct cagcagatga
     2401 taatatcaag atacctgctg agcatctgcg ggttccgctt ccaccgtcag cggatgataa
     2461 tctcaagaca ccttccgagc gtcagctcac tccccttcca ccctcagctc caccctcagc
     2521 agatgataat atcaagacac ctgccgagcg tctgcggggg ccgcttccac cctcagccga
     2581 tgataatctc aagacacctt ccgagcgtca gctcactccc cttccaccct cagctccacc
     2641 ctcagcagat gataatatca agacacctgc cgagcgtctg cgggggccgc ttccaccctc
     2701 agccgatgat aatctcaaga caccttccga gcgtcagctc actccccttc caccctcagc
     2761 tccaccctca gcagatgata atatcaagac acctgccgag cgtctgcggg ggccgcttcc
     2821 accctcagcc gatgataatc tcaagacacc ttccgagcgt cagctcactc cccttccacc
     2881 ctcagctcca ccctcagcag atgataatat caagacacct gccgagcgtc tgcgggggcc
     2941 gcttccaccc tcagccgatg ataatctcaa gacaccttcc gagcgtcagc tcactcccct
     3001 tccaccctca gctccaccct cagcagatga taatatcaag acacctgccg agcgtctgcg
     3061 ggggccgctt ccaccctcag ccgatgataa tctcaagaca ccttccgagc gtcagctcac
     3121 tccccttcca ccctcagctc caccctcagc agatgataat atcaagacac ctgccgagcg
     3181 tctgcgggga gcgtctgtgg cgtctgcggg ggccgcttcc accctcagcc gatgataatc
     3241 tcaagacacc ttccaaacgt cagctcactc cccttccacc ctcagctcca ccctcagcag
     3301 atgataatat caagacacct gccgagcgtc tgcgggggcc gcttccaccc tcagccgatg
     3361 ataatctcaa gacaccttcc gagcgtcagc tcactcccct tccaccctca gctccaccct
     3421 cagcagatga taatatcaag acacctgccg agcgtctgcg ggggccgctt ccaccctcag
     3481 cggatgataa tctcaagaca ccttccgagc gtcagctcac tccccttcca ccctcagctc
     3541 caacctcagc agatgataat atcaagacac ctgccgagcg tctgcggggg ccgcttccac
     3601 cctcagccga tgataatctc aagacacctc ccttagctac tcaggaggct gaggcagaaa
     3661 aaccacgcaa acccaagagg cagagggcgg ctgagatgga accacctccc gaacccaaga
     3721 ggcggagggt cggtgacgtg gaaccgtcac gcaaacccaa gaggcggagg gccgctgacg
     3781 tggaaccatc atcacccgaa cccaagaggc ggagggtcgg tgatgtggaa ccgtcacgca
     3841 aacccaagag gcggagggcc gctgacgtgg aaccatcatc acccgaaccc aagaggcgga
     3901 ggttgagcga ttacaaggat gacgacgata agtgataaac ccgctgatca gcctcgactg
     3961 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4021 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4081 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4141 aagacaatag caggcatgcc aattggaaga ctaatgcgtt ttcgagatat ctagacccag
     4201 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     4261 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgcagct ctggcccgtg
     4321 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     4381 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     4441 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     4501 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     4561 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     4621 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     4681 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     4741 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtc cctgcgccgg
     4801 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     4861 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     4921 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     4981 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     5041 ttaggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat
     5101 atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc
     5161 gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt
     5221 gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag
     5281 atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag
     5341 agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag
     5401 caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa
     5461 ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct
     5521 acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg
     5581 ctgcgtaaca tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
     5641 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt
     5701 cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc
     5761 tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt
     5821 atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg
     5881 gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct
     5941 ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg
     6001 atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg ccggtggtgc
     6061 tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg
     6121 cccaggactc tagctatagt tctagtggtt ggctactaat aggttgtatt gatgttggac
     6181 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     6241 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     6301 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     6361 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     6421 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6481 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6541 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6601 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6721 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6781 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6841 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6901 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6961 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7021 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7081 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7141 tgcgttatcc cctgattctg tggataaccg tattaccgct agcatggatc tcggggacgt
     7201 ctaactacta agcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     7261 gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     7321 aatccgccgg gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga
     7381 cgcccgccat aaactgccag gcatcaaact aagcagaagg ccatc
//
