LOCUS       sequence-375652-        7128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-TTI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7128)
  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 7128)
  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-TTI1
FEATURES             Location/Qualifiers
     source          1..7128
                     /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(3898..3997)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4109..4128)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4187..4206)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4780..5313)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5502..5507,5508..5524,5525..5530))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      6187..6775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6676..6695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      6940..7026
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(7118..7128,1..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 gcatggcagt
      601 ttttgatact cctgaggagg cctttggtgt cttacgtcca gtctgtgttc agctcacaaa
      661 gacccagaca gtggagaatg tggagcatct gcagacacga ctacaagctg tgagtgacag
      721 tgcccttcag gaacttcagc agtacatcct cttccctctg cgatttaccc tgaagacccc
      781 aggtcccaaa agagagcgtt tgatccaaag tgtggtggaa tgcctcacat ttgtcctttc
      841 ttcaacatgt gtgaaagaac aggagcttct ccaggaactc ttttcagaac tctctgcttg
      901 tctgtattca cccagctccc aaaaacctgc ggctgtgtcc gaggagttga aattggctgt
      961 gatccaggga cttagcacat taatgcactc agcttatggg gacatcattc tgacttttta
     1021 tgagccctcc attctgccac gtttaggatt tgctgtatct ttactgttag gccttgcaga
     1081 acaggagaaa tcaaagcaaa ttaaaattgc tgccttaaaa tgtttacagg ttctactctt
     1141 gcagtgtgat tgtcaggacc atccaaggtc attggatgaa cttgaacaaa agcagctggg
     1201 ggatttgttt gcctcttttt tacctggaat ctcaactgca ctgaccaggc ttatcacagg
     1261 agactttaaa caaggtcaca gcattgtcgt atcttcccta aagatctttt acaagacagt
     1321 gagcttcatt atggctgatg aacagctcaa aagaatctca aaggtccaag caaaacctgc
     1381 agttgagcac agagtagcag agctgatggt ttacagggaa gcagattggg taaaaaagac
     1441 tggcgacaag ttgactatcc ttattaaaaa gataattgag tgtgtttctg ttcacccaca
     1501 ctggaaggtg agactggaac tggtagaact tgtggaggac cttcttttga agtgcagtca
     1561 atcattggtc gaatgtgctg gtccccttct gaaggcctta gtgggactag taaatgatga
     1621 gagtcctgaa atccaagccc agtgcaataa agttctgaga cattttgcag atcaaaaagt
     1681 agtggtgggc aacaaagccc tcgctgacat cttgtcagaa agcctgcatt cccttgccac
     1741 atctcttcct cgcctaatga actcccaaga tgaccagggc aaattctcta ctctttcctt
     1801 gttacttggt tatctgaaac tcttgggccc aaaaataaac tttgtcctca actctgtggc
     1861 ccatctccag cggctttcca aagcactcat ccaagttcta gagctagacg tggctgacat
     1921 caagattgtt gaggaacggc gttggaactc tgatgatctg aatgcttctc caaagacctc
     1981 agccacacag ccttggaacc gcatccagag gagatatttc cgcttcttca ctgatgagag
     2041 aatcttcatg ctcttgaggc aggtttgtca gctacttggt tattatggga atctttattt
     2101 gcttgtggat cactttatgg aactttacca tcaatctgtg gtttaccgga agcaagctgc
     2161 catgatcctt aatgaactgg ttacaggggc tgctgggctg gaggttgagg atcttcacga
     2221 aaaacatatt aaaacaaacc cagaagaact gagagagatt gtgacatcta tacttgaaga
     2281 atacacaagt caagaaaatt ggtatttggt tacctgtctt gaaactgagg aaatgggaga
     2341 ggagctgatg atggagcacc caggcctcca agccatcacg tctggtgaac acacctgcca
     2401 agttacatct tttctagcct tctcaaagcc aagtcccact atttgctcca tgaacagtaa
     2461 catctggcaa atatgcattc agttggaagg aattggccag tttgcatatg cactaggaaa
     2521 agacttctgt ttgctcttga tgtcagccct ttatccagta ctggagaagg ctggagacca
     2581 aaccctactc attagtcagg tggctaccag caccatgatg gacgtttgcc gtgcttgtgg
     2641 ctacgactcc ctgcagcacc tgatcaatca aaattcagac tatttagtga atgggatctc
     2701 tttaaatctg cgtcatctgg ctctgcatcc tcatacccca aaggtcctgg aagtcatgct
     2761 gcggaactca gatgctaacc tgcttccttt ggtggcagat gtggttcaag atgtcttggc
     2821 caccctggac caattttacg ataagagagc tgcttccttt gtcagcgttc tgcatgctct
     2881 gatggcagca ttagcccagt ggttcccaga cacaggtaat cttgggcacc tccaagagca
     2941 aagtttagga gaagagggaa gtcatttgaa ccaaagacca gcagctcttg agaagagcac
     3001 caccacagct gaagacatcg aacagttttt gctgaactac ctcaaagaga aggatgtggc
     3061 agatggaaat gtctcggatt ttgataatga agaagaggaa cagtcagtcc ctcccaaagt
     3121 ggatgagaat gacacccgtc cagatgtgga gccaccactg ccattgcaga tccaaatagc
     3181 catggacgtg atggaacgct gcatccactt gttgtcagat aaaaatctgc aaatccgcct
     3241 gaaggtcttg gatgtgctgg atctgtgtgt ggttgttctt cagtcccaca aaaaccagct
     3301 gcttcccttg gctcatcagg cctggccctc gctcgttcac cgactcacac gggacgcccc
     3361 cctggcagtg cttagagcct tcaaggtttt acgtaccctg ggaagcaagt gtggtgactt
     3421 tcttcgcagc cggttctgca aagatgtcct gccaaagctg gctggctccc tagtcaccca
     3481 ggcccccatc agtgccaggg ctggaccagt ttactcgcac acgctggcct tcaagttgca
     3541 gctggctgtc ttacagggcc tgggccccct ctgtgagaga ctggacctag gtgagggtga
     3601 cctgaataaa gtggctgatg cctgcttgat ttacctcagt gtcaaacagc ccgtgaaatt
     3661 acaagaggct gccaggagcg tcttcctcca cttgatgaag gtggacccag actccacctg
     3721 gttcctcctg aacgagcttt actgccccgt gcagttcaca cctccccacc ccagcctcca
     3781 ccctgtgcag ctgcacgggg ccagcgggca gcagaacccc tacacgacca acgtgctcca
     3841 gctgctcaag gagctgcagt gacaattgga agactaatgc gttttcgaga tatctagacc
     3901 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     3961 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gctctggccc
     4021 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     4081 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     4141 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     4201 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     4261 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     4321 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     4381 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     4441 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gtccctgcgc
     4501 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     4561 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     4621 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     4681 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     4741 aaattaggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc
     4801 gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg
     4861 atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg
     4921 cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc
     4981 gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc
     5041 gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg
     5101 gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc
     5161 gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag
     5221 cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc
     5281 ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc
     5341 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac
     5401 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     5461 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
     5521 cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
     5581 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
     5641 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
     5701 cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg
     5761 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
     5821 tcgcccagga ctctagctat agttctagtg gttggctact aataggttgt attgatgttg
     5881 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     5941 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     6001 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     6061 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     6121 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6181 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6241 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6301 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6361 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6421 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6481 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6541 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6601 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6661 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6721 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6781 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6841 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gctagcatgg atctcgggga
     6901 cgtctaacta ctaagcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     6961 tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     7021 acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca
     7081 ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatc
//
