LOCUS       sequence-375651-        5688 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-XRCC6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5688)
  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 5688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-XRCC6
FEATURES             Location/Qualifiers
     source          1..5688
                     /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(2458..2557)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2669..2688)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2747..2766)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(3340..3873)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4062..4067,4068..4084,4085..4090))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4747..5335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5236..5255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5500..5586
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(5678..5688,1..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 gtatgtcagg
      601 gtgggagtca tattacaaaa ccgagggcga tgaagaagca gaggaagaac aagaagagaa
      661 ccttgaagca agtggagact ataaatattc aggaagagat agtttgattt ttttggttga
      721 tgcctccaag gctatgtttg aatctcagag tgaagatgag ttgacacctt ttgacatgag
      781 catccagtgt atccaaagtg tgtacatcag taagatcata agcagtgatc gagatctctt
      841 ggctgtggtg ttctatggta ccgagaaaga caaaaattca gtgaatttta aaaatattta
      901 cgtcttacag gagctggata atccaggtgc aaaacgaatt ctagagcttg accagtttaa
      961 ggggcagcag ggacaaaaac gtttccaaga catgatgggc cacggatctg actactcact
     1021 cagtgaagtg ctgtgggtct gtgccaacct ctttagtgat gtccaattca agatgagtca
     1081 taagaggatc atgctgttca ccaatgaaga caacccccat ggcaatgaca gtgccaaagc
     1141 cagccgggcc aggaccaaag ccggtgatct ccgagataca ggcatcttcc ttgacttgat
     1201 gcacctgaag aaacctgggg gctttgacat atccttgttc tacagagata tcatcagcat
     1261 agcagaggat gaggacctca gggttcactt tgaggaatcc agcaagctag aagacctgtt
     1321 gcggaaggtt cgcgccaagg agacaaggaa gcgagcactc agcaggttaa agctgaagct
     1381 caacaaagat atagtgatct ctgtgggcat ttataatctg gtccagaagg ctctcaagcc
     1441 tcctccaata aagctctatc gggaaacaaa tgaaccagtg aaaaccaaga cccggacctt
     1501 taatacaagt acaggcggtt tgcttctgcc tagcgatacc aagaggtctc agatctatgg
     1561 gagtcgtcag attatactgg agaaagagga aacagaagag ctaaaacggt ttgatgatcc
     1621 aggtttgatg ctcatgggtt tcaagccgtt ggtactgctg aagaaacacc attacctgag
     1681 gccctccctg ttcgtgtacc cagaggagtc gctggtgatt gggagctcaa ccctgttcag
     1741 tgctctgctc atcaagtgtc tggagaagga ggttgcagca ttgtgcagat acacaccccg
     1801 caggaacatc cctccttatt ttgtggcttt ggtgccacag gaagaagagt tggatgacca
     1861 gaaaattcag gtgactcctc caggcttcca gctggtcttt ttaccctttg ctgatgataa
     1921 aaggaagatg ccctttactg aaaaaatcat ggcaactcca gagcaggtgg gcaagatgaa
     1981 ggctatcgtt gagaagcttc gcttcacata cagaagtgac agctttgaga accccgtgct
     2041 gcagcagcac ttcaggaacc tggaggcctt ggccttggat ttgatggagc cggaacaagc
     2101 agtggacctg acattgccca aggttgaagc aatgaataaa agactgggct ccttggtgga
     2161 tgagtttaag gagcttgttt acccaccaga ttacaatcct gaagggaaag ttaccaagag
     2221 aaaacacgat aatgaaggtt ctggaagcaa aaggcccaag gtggagtatt cagaagagga
     2281 gctgaagacc cacatcagca agggtacgct gggcaagttc actgtgccca tgctgaaaga
     2341 ggcctgccgg gcttacgggc tgaagagtgg gctgaagaag caggagctgc tggaagccct
     2401 caccaagcac ttccaggact gacaattgga agactaatgc gttttcgaga tatctagacc
     2461 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2521 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gctctggccc
     2581 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     2641 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     2701 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     2761 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2821 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2881 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2941 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3001 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gtccctgcgc
     3061 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     3121 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3181 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3241 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3301 aaattaggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc
     3361 gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg
     3421 atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg
     3481 cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc
     3541 gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc
     3601 gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg
     3661 gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc
     3721 gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag
     3781 cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc
     3841 ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc
     3901 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac
     3961 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     4021 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
     4081 cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
     4141 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
     4201 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
     4261 cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg
     4321 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
     4381 tcgcccagga ctctagctat agttctagtg gttggctact aataggttgt attgatgttg
     4441 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     4501 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     4561 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     4621 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     4681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4741 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4801 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4861 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4921 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4981 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5041 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5101 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5161 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5221 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5281 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5341 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5401 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gctagcatgg atctcgggga
     5461 cgtctaacta ctaagcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     5521 tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     5581 acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca
     5641 ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatc
//
