LOCUS       sequence-375654-        3924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3924)
  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 3924)
  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-Linker
FEATURES             Location/Qualifiers
     source          1..3924
                     /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(694..793)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(905..924)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(983..1002)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1576..2109)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2298..2303,2304..2320,2321..2326))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      2983..3571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3472..3491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3736..3822
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(3914..3924,1..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 gtggtaccac
      601 cggtgaattc cacgtgaagc ttccatggct cgagcccggg tacgtaacgc gtgcatgcca
      661 attggaagac taatgcgttt tcgagatatc tagacccagc tttcttgtac aaagttggca
      721 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
      781 aaatcattat ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac
      841 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
      901 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
      961 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1021 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1081 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1141 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1201 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1261 gaaaatattg ttgatgcgct ggcagtgtcc ctgcgccggt tgcattcgat tcctgtttgt
     1321 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     1381 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     1441 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     1501 gatttctcac ttgataacct tatttttgac gaggggaaat taggccggga agccgatctc
     1561 ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc
     1621 ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac
     1681 gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt
     1741 ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact
     1801 acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg
     1861 gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga
     1921 gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag
     1981 agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat
     2041 acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     2101 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     2161 gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg
     2221 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     2281 tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct
     2341 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat
     2401 ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt caggcattgg
     2461 cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga
     2521 tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat gaagtggttc
     2581 gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt
     2641 ctagtggttg gctactaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2701 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2761 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2821 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     2881 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttttcgtt
     2941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3121 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3601 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3661 ggataaccgt attaccgcta gcatggatct cggggacgtc taactactaa gcgagagtag
     3721 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt
     3781 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     3841 aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     3901 catcaaacta agcagaaggc catc
//
