LOCUS       sequence-375632-        4531 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway entry vector for an inducible 3xFLAG-BCAS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4531)
  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 4531)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-BCAS2
FEATURES             Location/Qualifiers
     source          1..4531
                     /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(1301..1400)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1512..1531)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1590..1609)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2183..2716)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2905..2910,2911..2927,2928..2933))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3590..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4079..4098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4343..4429
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4521..4531,1..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 gcgggcacag
      601 gtttggtggc tggagaggtt gtggtggatg cgctgccgta ttttgatcaa ggttatgaag
      661 cccctggtgt gcgggaagcg gctgcagcgc tggtggagga ggaaactcgc agataccgac
      721 ctactaagaa ctacctgagc tacctgacag ccccggatta ttctgccttt gaaactgaca
      781 taatgagaaa tgaatttgaa agactggctg ctcgacaacc aattgaattg ctcagtatga
      841 aacgatatga gcttccagcc ccctcctctg gtcaaaaaaa tgacattact gcatggcaag
      901 aatgtgtaaa caattctatg gcccagttag agcatcaagc agttagaatt gagaatctgg
      961 aactaatgtc acagcatgga tgtaatgcct ggaaagtata caatgaaaat ctagttcata
     1021 tgattgaaca cgcacagaag gaacttcaga agttaagaaa acatattcaa gatttaaact
     1081 ggcagagaaa gaacatgcaa ctcacagctg gatctaaatt gagagaaatg gagtcaaatt
     1141 gggtatccct ggtcagtaag aattatgaga ttgaacggac tattgttcag ctagaaaatg
     1201 aaatctatca aattaagcag caacatggag aggcaaacaa agaaaacatc cggcaagact
     1261 tctgagaatt ggaagactaa tgcgttttcg agatatctag acccagcttt cttgtacaaa
     1321 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1381 caaaataaaa tcattatttg ccatccagct gcagctctgg cccgtgtctc aaaatctctg
     1441 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1501 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1561 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1621 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1681 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1741 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1801 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1861 ttcaggtgaa aatattgttg atgcgctggc agtgtccctg cgccggttgc attcgattcc
     1921 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     1981 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2041 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2101 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattag gccgggaagc
     2161 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     2221 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     2281 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     2341 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     2401 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     2461 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     2521 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     2581 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     2641 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     2701 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     2761 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     2821 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     2881 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     2941 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3001 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3061 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3121 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3181 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3241 tatagttcta gtggttggct actaataggt tgtattgatg ttggacgagt cggaatcgca
     3301 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     3361 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     3421 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     3481 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     3541 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4201 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4261 attctgtgga taaccgtatt accgctagca tggatctcgg ggacgtctaa ctactaagcg
     4321 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt
     4381 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     4441 ggatttgaac gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     4501 tgccaggcat caaactaagc agaaggccat c
//
