LOCUS       sequence-375751-        4276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-PFN2_variant2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4276)
  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 4276)
  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-PFN2_variant2
FEATURES             Location/Qualifiers
     source          1..4276
                     /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             522..587
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1046..1145)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1257..1276)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1335..1354)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1928..2461)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2650..2655,2656..2672,2673..2678))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3335..3923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3824..3843
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4088..4174
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4266..4276,1..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 ggactacaaa gaccatgacg
      541 gtgattataa agatcatgac atcgattaca aggatgacga tgacaagatg gccggttggc
      601 agagctacgt ggataacctg atgtgcgatg gctgctgcca ggaggccgcc attgtcggct
      661 actgcgacgc caaatacgtc tgggcagcca cggcaggggg cgtctttcag agcattacgc
      721 caatagaaat agatatgatt gtaggaaaag accgggaagg tttctttacc aacggtttgg
      781 ctcttggcgc gaagaaatgc tcagtgatca gagatagtct atacgtcgat ggtgactgca
      841 caatggacat ccggacaaag agtcaaggtg gggagccaac atacaatgtg gctgtcggta
      901 gagctggtag agtcttggtc tttgtaatgg gaaaagaagg ggtccatgga ggcggattga
      961 ataagaaggc atactcaatg gcaaaatact tgagagactc tgggttctag caattggaag
     1021 actaatgcgt tttcgagata tctagaccca gctttcttgt acaaagttgg cattataaga
     1081 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1141 atttgccatc cagctgcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1201 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1261 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1321 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1381 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1441 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1501 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1561 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1621 tgttgatgcg ctggcagtgt ccctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1681 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     1741 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1801 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1861 acttgataac cttatttttg acgaggggaa attaggccgg gaagccgatc tcggcttgaa
     1921 cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc
     1981 ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca
     2041 cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc
     2101 cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct
     2161 gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg
     2221 cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct
     2281 gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc
     2341 gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc
     2401 cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca
     2461 tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat
     2521 gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg
     2581 ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac
     2641 ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt
     2701 ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc
     2761 tggctggcga acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg
     2821 ctgttcttct acggcaaggt gctgtgcacg gatctgccct ggcttcagga gatcggaaga
     2881 cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc
     2941 ggttttctgg aaggcgagca tcgtttgttc gcccaggact ctagctatag ttctagtggt
     3001 tggctactaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3061 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3121 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3181 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     3241 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3301 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3421 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3481 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3541 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3601 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3661 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3721 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3781 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3841 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3901 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3961 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4021 gtattaccgc tagcatggat ctcggggacg tctaactact aagcgagagt agggaactgc
     4081 caggcatcaa ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt ttatctgttg
     4141 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgt
     4201 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaac
     4261 taagcagaag gccatc
//
