LOCUS       sequence-272308-        5050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used as a donor vector to clone into pSLIK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5050)
  AUTHORS   Pereira EJ, Burns JS, Lee CY, Marohl T, Calderon D, Wang L, Atkins 
            KA, Wang CC, Janes KA
  TITLE     Sporadic activation of an oxidative stress-dependent NRF2-p53 
            signaling network in breast epithelial spheroids and 
            premalignancies.
  JOURNAL   Sci Signal. 2020 Apr 14;13(627). pii: 13/627/eaba4200. doi: 
            10.1126/scisignal.aba4200.
  PUBMED    32291314
REFERENCE   2  (bases 1 to 5050)
  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: "Sci 
            Signal."; date: "2020-04-14"; volume: "13(627). pii: 
            13/627/eaba4200. doi"; pages: " 10.1126/scisignal.aba4200"
COMMENT     Sequence Label: pEN_TTmiRc2_p53(R280K)_V5
FEATURES             Location/Qualifiers
     source          1..5050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..27
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             37..1215
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLML
                     SPDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQ
                     GSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIY
                     KQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEP
                     PEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGKD
                     RRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEM
                     FRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     CDS             1249..1290
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(1338..1437)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1549..1568)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1627..1646)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2220..2753)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2942..2947,2948..2964,2965..2970))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3627..4215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4116..4135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4380..4466
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4558..4585
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4593..4616
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    4635..4734
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4766..5050,1..30)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4774..4792
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4810..4828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4845..4863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4881..4899
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4917..4935
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4952..4970
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4988..5006
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 agagctcgtt tagtgaaccg tcagatcgcc actagtatgg aggagccgca gtcagatcct
       61 agcgtcgagc cccctctgag tcaggaaaca ttttcagacc tatggaaact acttcctgaa
      121 aacaacgttc tgtccccctt gccgtcccaa gcaatggatg atttgatgct gtccccggac
      181 gatattgaac aatggttcac tgaagaccca ggtccagatg aagctcccag aatgccagag
      241 gctgctcccc gcgtggcccc tgcaccagca gctcctacac cggcggcccc tgcaccagcc
      301 ccctcctggc ccctgtcatc ttctgtccct tcccagaaaa cctaccaggg cagctacggt
      361 ttccgtctgg gcttcttgca ttctgggaca gccaagtctg tgacttgcac gtactcccct
      421 gccctcaaca agatgttttg ccaactggcc aagacctgcc ctgtgcagct gtgggttgat
      481 tccacacccc cgcccggcac ccgcgtccgc gccatggcca tctacaagca gtcacagcac
      541 atgacggagg ttgtgaggcg ctgcccccac catgagcgct gctcagatag cgatggtctg
      601 gcccctcctc agcatcttat ccgagtggaa ggaaatttgc gtgtggagta tttggatgac
      661 agaaacactt ttcgacatag tgtggtggtg ccctatgagc cgcctgaggt tggctctgac
      721 tgtaccacca tccactacaa ctacatgtgt aacagttcct gcatgggcgg catgaaccgg
      781 aggcccatcc tcaccatcat cacactggaa gactccagtg gtaatctact gggacggaac
      841 agctttgagg tgcgtgtttg tgcctgtcct gggaaagacc ggcgcacaga ggaagagaat
      901 ctccgcaaga aaggggagcc tcaccacgag ctgcccccag ggagcactaa gcgagcactg
      961 cccaacaaca ccagctcctc tccccagcca aagaagaaac cactggatgg agaatatttc
     1021 acccttcaga tccgtgggcg tgagcgcttc gagatgttcc gagagctgaa tgaggccttg
     1081 gaactcaagg atgcccaggc tgggaaggag ccagggggga gcagggctca ctccagccac
     1141 ctgaagtcca aaaagggtca gtctacctcc cgccataaaa aactcatgtt caagacagaa
     1201 gggcctgact cagacaaggg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc
     1261 cctaaccctc tcctcggtct cgattctacg cgtaccggtt agcaattgga agactaatgc
     1321 gttttcgaga tatctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1381 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1441 tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1501 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1561 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1621 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1681 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1741 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1801 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1861 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1921 cgctggcagt gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1981 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2041 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2101 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2161 accttatttt tgacgagggg aaattaggcc gggaagccga tctcggcttg aacgaattgt
     2221 taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt
     2281 gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca
     2341 ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc
     2401 cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac
     2461 ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc
     2521 gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg
     2581 gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat
     2641 ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac
     2701 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa catcgttgct
     2761 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     2821 catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc
     2881 accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta
     2941 cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg
     3001 tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt cctggctggc
     3061 gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct tgctgttctt
     3121 ctacggcaag gtgctgtgca cggatctgcc ctggcttcag gagatcggaa gacctcggcc
     3181 gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc tcggttttct
     3241 ggaaggcgag catcgtttgt tcgcccagga ctctagctat agttctagtg gttggctact
     3301 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3361 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3421 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3481 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3541 cggcgcaagc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3601 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3661 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3721 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3781 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3841 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3901 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3961 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4021 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4081 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4141 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4201 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4261 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4321 gctagcatgg atctcgggga cgtctaacta ctaagcgaga gtagggaact gccaggcatc
     4381 aaataaaacg aaaggctcag tcggaagact gggcctttcg ttttatctgt tgtttgtcgg
     4441 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac
     4501 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa actaagcaga
     4561 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctgtt agttagttac
     4621 ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt tcgttgcaac
     4681 aaattgataa gcaatgcttt tttataatgc caactttgta caaaaaagca ggctttaaag
     4741 gaaccaattc agtcgactgg atcccgagtt tactccctat cagtgataga gaacgtatgt
     4801 cgagtttact ccctatcagt gatagagaac gatgtcgagt ttactcccta tcagtgatag
     4861 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc
     4921 tatcagtgat agagaacgta tgtcgagttt atccctatca gtgatagaga acgtatgtcg
     4981 agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggaggc
     5041 ctatataagc
//
