LOCUS       sequence-375719-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-KIF20A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6526)
  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 6526)
  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-KIF20A
FEATURES             Location/Qualifiers
     source          1..6526
                     /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(3296..3395)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3507..3526)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3585..3604)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4178..4711)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4900..4905,4906..4922,4923..4928))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5585..6173
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6074..6093
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      6338..6424
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(6516..6526,1..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 tcgcaaggga
      601 tcctttctcc gccagcgggc ttgctgtccg atgacgatgt cgtagtttct cccatgtttg
      661 agtccacagc tgcagatttg gggtctgtgg tacgcaagaa cctgctatca gactgctctg
      721 tcgtctctac ctccctagag gacaagcagc aggttccatc tgaggacagt atggagaagg
      781 tgaaagtata cttgagggtt aggcccttgt taccttcaga gttggaacga caggaagatc
      841 agggttgtgt ccgtattgag aatgtggaga cccttgttct acaagcaccc aaggactctt
      901 ttgccctgaa gagcaatgaa cggggaattg gccaagccac acacaggttc accttttccc
      961 agatctttgg gccagaagtg ggacaggcat ccttcttcaa cctaactgtg aaggagatgg
     1021 taaaggatgt actcaaaggg cagaactggc tcatctatac atatggagtc actaactcag
     1081 ggaaaaccca cacgattcaa ggtaccatca aggatggagg gattctcccc cggtccctgg
     1141 cgctgatctt caatagcctc caaggccaac ttcatccaac acctgatctg aagcccttgc
     1201 tctccaatga ggtaatctgg ctagacagca agcagatccg acaggaggaa atgaagaagc
     1261 tgtccctgct aaatggaggc ctccaagagg aggagctgtc cacttccttg aagaggagtg
     1321 tctacatcga aagtcggata ggtaccagca ccagcttcga cagtggcatt gctgggctct
     1381 cttctatcag tcagtgtacc agcagtagcc agctggatga aacaagtcat cgatgggcac
     1441 agccagacac tgccccacta cctgtcccgg caaacattcg cttctccatc tggatctcat
     1501 tctttgagat ctacaacgaa ctgctttatg acctattaga accgcctagc caacagcgca
     1561 agaggcagac tttgcggcta tgcgaggatc aaaatggcaa tccctatgtg aaagatctca
     1621 actggattca tgtgcaagat gctgaggagg cctggaagct cctaaaagtg ggtcgtaaga
     1681 accagagctt tgccagcacc cacctcaacc agaactccag ccgcagtcac agcatcttct
     1741 caatcaggat cctacacctt cagggggaag gagatatagt ccccaagatc agcgagctgt
     1801 cactctgtga tctggctggc tcagagcgct gcaaagatca gaagagtggt gaacggttga
     1861 aggaagcagg aaacattaac acctctctac acaccctggg ccgctgtatt gctgcccttc
     1921 gtcaaaacca gcagaaccgg tcaaagcaga acctggttcc cttccgtgac agcaagttga
     1981 ctcgagtgtt ccaaggtttc ttcacaggcc gaggccgttc ctgcatgatt gtcaatgtga
     2041 atccctgtgc atctacctat gatgaaactc ttcatgtggc caagttctca gccattgcta
     2101 gccagcttgt gcatgcccca cctatgcaac tgggattccc atccctgcac tcgttcatca
     2161 aggaacatag tcttcaggta tcccccagct tagagaaagg ggctaaggca gacacaggcc
     2221 ttgatgatga tattgaaaat gaagctgaca tctccatgta tggcaaagag gagctcctac
     2281 aagttgtgga agccatgaag acactgcttt tgaaggaacg acaggaaaag ctacagctgg
     2341 agatgcatct ccgagatgaa atttgcaatg agatggtaga acagatgcaa cagcgggaac
     2401 agtggtgcag tgaacatttg gacacccaaa aggaactatt ggaggaaatg tatgaagaaa
     2461 aactaaatat cctcaaggag tcactgacaa gtttttacca agaagagatt caggagcggg
     2521 atgaaaagat tgaagagcta gaagctctct tgcaggaagc cagacaacag tcagtggccc
     2581 atcagcaatc agggtctgaa ttggccctac ggcggtcaca aaggttggca gcttctgcct
     2641 ccacccagca gcttcaggag gttaaagcta aattacagca gtgcaaagca gagctaaact
     2701 ctaccactga agagttgcat aagtatcaga aaatgttaga accaccaccc tcagccaagc
     2761 ccttcaccat tgatgtggac aagaagttag aagagggcca gaagaatata aggctgttgc
     2821 ggacagagct tcagaaactt ggtgagtctc tccaatcagc agagagagct tgttgccaca
     2881 gcactggggc aggaaaactt cgtcaagcct tgaccacttg tgatgacatc ttaatcaaac
     2941 aggaccagac tctggctgaa ctgcagaaca acatggtgct agtgaaactg gaccttcgga
     3001 agaaggcagc atgtattgct gagcagtatc atactgtgtt gaaactccaa ggccaggttt
     3061 ctgccaaaaa gcgccttggt accaaccagg aaaatcagca accaaaccaa caaccaccag
     3121 ggaagaaacc attccttcga aatttacttc cccgaacacc aacctgccaa agctcaacag
     3181 actgcagccc ttatgcccgg atcctacgct cacggcgttc ccctttactc aaatctgggc
     3241 cttttggcaa aaagtactaa caattggaag actaatgcgt tttcgagata tctagaccca
     3301 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     3361 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagc tctggcccgt
     3421 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     3481 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     3541 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3601 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3661 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3721 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3781 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3841 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt ccctgcgccg
     3901 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3961 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     4021 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     4081 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     4141 attaggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
     4201 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
     4261 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
     4321 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga
     4381 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
     4441 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
     4501 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
     4561 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
     4621 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
     4681 gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
     4741 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag
     4801 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
     4861 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct
     4921 tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
     4981 ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc
     5041 tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg
     5101 gatctgccct ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccggtggtg
     5161 ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc
     5221 gcccaggact ctagctatag ttctagtggt tggctactaa taggttgtat tgatgttgga
     5281 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     5341 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     5401 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     5461 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     5521 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5581 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5641 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5701 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5761 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5821 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5881 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5941 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6001 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6061 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6121 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6181 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6241 ctgcgttatc ccctgattct gtggataacc gtattaccgc tagcatggat ctcggggacg
     6301 tctaactact aagcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     6361 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     6421 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
     6481 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatc
//
