LOCUS       sequence-388102-        7738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-BirA* 3xFLAG-CSE1L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7738)
  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 7738)
  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 BirA*-3xFLAG-CSE1L
FEATURES             Location/Qualifiers
     source          1..7738
                     /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             join(519..869,870..872,873..1481)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1491..1556
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(4508..4607)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4719..4738)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4797..4816)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(5390..5923)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6112..6117,6118..6134,6135..6140))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      6797..7385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7286..7305
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      7550..7636
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(7728..7738,1..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 ccactagaat gaaggacaac accgtgcccc
      541 tgaagctgat cgccctgctg gccaacggcg agttccactc tggcgagcag ctgggagaga
      601 ccctgggaat gagcagagcc gccatcaaca agcacatcca gacactgaga gactggggag
      661 tggacgtgtt caccgtgcct ggcaagggct acagcctgcc tgagcctatc cagctgctga
      721 acgccaagca gatcctggga cagctggatg gcggaagcgt ggccgtgctg cctgtgatcg
      781 actccaccaa tcagtacctg ctggacagaa tcggagagct gaagtccggc gacgcctgca
      841 tcgccgagta ccagcaggct ggcagaggag gcagaggacg gaagtggttc agcccattcg
      901 gagccaacct gtacctgtcc atgttctgga gactggagca gggacctgct gctgccatcg
      961 gactgagtct ggtgatcgga atcgtgatgg ccgaggtgct gagaaagctg ggagccgaca
     1021 aggtgagagt gaagtggcct aatgacctgt acctccagga ccgcaagctg gctggcatcc
     1081 tggtggagct gacaggcaag acaggcgatg ccgctcagat cgtgatcgga gccggaatca
     1141 acatggccat gagaagagtg gaggagagcg tggtgaacca gggctggatc accctgcagg
     1201 aggctggcat caacctggac cggaacaccc tggccgccat gctgatcaga gagctgagag
     1261 ccgctctgga gctgttcgag caggagggac tggctcctta cctgagcaga tgggagaagc
     1321 tggacaactt catcaacaga cctgtgaagc tgatcatcgg cgacaaggaa atcttcggca
     1381 tctccagagg aatcgacaag cagggagctc tgctgctgga gcaggacgga atcatcaagc
     1441 cctggatggg cggagaaatc tccctgagaa gcgcagagaa gactagaatg gactacaaag
     1501 accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagatgg
     1561 aactcagcga tgcaaatctg caaacactaa cagaatattt aaagaaaaca cttgatcctg
     1621 atcctgccat ccgacgtcca gctgagaaat ttcttgaatc tgttgaagga aatcagaatt
     1681 atccactgtt gcttttgaca ttactggaga agtcccagga taatgttatc aaagtatgtg
     1741 cttcagtaac attcaaaaac tatattaaaa ggaactggag aattgttgaa gatgaaccaa
     1801 acaaaatttg tgaagccgat cgagtggcca ttaaagccaa catagtgcac ttgatgctta
     1861 gcagcccaga gcaaattcag aagcagttaa gtgatgcaat tagcattatt ggcagagaag
     1921 attttccaca gaaatggcct gacttgctga cagaaatggt gaatcgcttt cagagtggag
     1981 atttccatgt tattaatgga gtcctccgta cagcacattc attatttaaa agataccgtc
     2041 atgaatttaa gtcaaacgag ttatggactg aaattaagct tgttctggat gcctttgctt
     2101 tgcctttgac taatcttttt aaggccacta ttgaactctg cagtacccat gcaaatgatg
     2161 cctctgccct gaggattctg ttttcttccc tgatcctgat ctcaaaattg ttctatagtt
     2221 taaactttca ggatctccct gaattttttg aagataatat ggaaacttgg atgaataatt
     2281 ttcatactct cttaacattg gataataagc ttttacaaac tgatgatgaa gaggaagccg
     2341 gcttattgga gctcttaaaa tcccagattt gtgataatgc cgcactctat gcacaaaagt
     2401 acgatgaaga attccagcga tacctgcctc gttttgttac agccatctgg aatttactag
     2461 ttacaacggg tcaagaggtt aaatatgatt tgttggtaag taatgcaatt caatttctgg
     2521 cttcagtttg tgagagacct cattataaga atctatttga ggaccagaac acgctgacaa
     2581 gtatctgtga aaaggttatt gtgcctaaca tggaatttag agctgctgat gaagaagcat
     2641 ttgaagataa ttctgaggag tacataagga gagatttgga aggatctgat attgatacta
     2701 gacgcagggc tgcttgtgat ctggtacgag gattatgcaa gttttttgag ggacctgtga
     2761 caggaatctt ctctggttat gttaattcca tgctgcagga atacgcaaaa aatccatctg
     2821 tcaactggaa acacaaagat gcagccatct acctagtgac atctttggca tcaaaagccc
     2881 aaacacagaa gcatggaatt acacaagcaa atgaacttgt aaacctaact gagttctttg
     2941 tgaatcacat cctccctgat ttaaaatcag ctaatgtgaa tgaatttcct gtccttaaag
     3001 ctgacggtat caaatatatt atgattttta gaaatcaagt gccaaaagaa catcttttag
     3061 tctcgattcc tctcttgatt aatcatcttc aagctgaaag tattgttgtt catacttacg
     3121 cagctcatgc tcttgaacgg ctctttacta tgcgagggcc taacaatgcc actctcttta
     3181 cagctgcaga aatcgcaccg tttgttgaga ttctgctaac aaaccttttc aaagctctca
     3241 cacttcctgg ctcttcagaa aatgaatata ttatgaaagc tatcatgaga agtttttctc
     3301 tcctacaaga agccataatc ccctacatcc ctactctcat cactcagctt acacagaagc
     3361 tattagctgt tagtaagaac ccaagcaaac ctcactttaa tcactacatg tttgaagcaa
     3421 tatgtttatc cataagaata acttgcaaag ctaaccctgc tgctgttgta aattttgagg
     3481 aggctttgtt tttggtgttt actgaaatct tacaaaatga tgtgcaagaa tttattccat
     3541 acgtctttca agtgatgtct ttgcttctgg aaacacacaa aaatgacatc ccgtcttcct
     3601 atatggcctt atttcctcat ctccttcagc cagtgctttg ggaaagaaca ggaaatattc
     3661 ctgctctagt gaggcttctt caagcattct tagaacgcgg ttcaaacaca atagcaagtg
     3721 ctgcagctga caaaattcct gggttactag gtgtctttca gaagctgatt gcatccaaag
     3781 caaatgacca ccaaggtttt tatcttctaa acagtataat agagcacatg cctcctgaat
     3841 cagttgacca atataggaaa caaatcttca ttctgctatt ccagagactt cagaattcca
     3901 aaacaaccaa gtttatcaag agttttttag tctttattaa tttgtattgc ataaaatatg
     3961 gggcactagc actacaagaa atatttgatg gtatacaacc aaaaatgttt ggaatggttt
     4021 tggaaaaaat tattattcct gaaattcaga aggtatctgg aaatgtagag aaaaagatct
     4081 gtgcggttgg cataaccaaa ttactaacag aatgtccccc aatgatggac actgagtata
     4141 ccaaactgtg gactccatta ttacagtctt tgattggtct ttttgagtta cccgaagatg
     4201 ataccattcc tgatgaggaa cattttattg acatagaaga tacaccagga tatcagactg
     4261 ccttctcaca gttggcattt gctgggaaaa aagagcatga tcctgtaggt caaatggtga
     4321 ataaccccaa aattcacctg gcacagtcac ttcacaagtt gtctaccgcc tgtccaggaa
     4381 gggttccatc aatggtgagc accagcctga atgcagaagc gctccagtat ctccaagggt
     4441 accttcaggc agccagtgtg acactgcttt aacaattgga agactaatgc gttttcgaga
     4501 tatctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     4561 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca
     4621 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     4681 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     4741 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     4801 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     4861 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     4921 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     4981 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     5041 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     5101 gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     5161 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     5221 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     5281 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     5341 tgacgagggg aaattaggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg
     5401 tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag
     5461 ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt
     5521 tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc
     5581 cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa
     5641 cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg
     5701 tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg
     5761 ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt
     5821 cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag
     5881 ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct gctccataac
     5941 atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt
     6001 acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     6061 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga
     6121 accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac
     6181 ccggcaacct tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc
     6241 aaggtttcgg tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag
     6301 gtgctgtgca cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc
     6361 ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag
     6421 catcgtttgt tcgcccagga ctctagctat agttctagtg gttggctact aataggttgt
     6481 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     6541 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     6601 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     6661 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     6721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7261 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7321 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7381 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7441 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gctagcatgg
     7501 atctcgggga cgtctaacta ctaagcgaga gtagggaact gccaggcatc aaataaaacg
     7561 aaaggctcag tcggaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     7621 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg
     7681 gtggcgggca ggacgcccgc cataaactgc caggcatcaa actaagcaga aggccatc
//
