LOCUS       sequence-272297-        4093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used as a donor vector containing N-term 3xFLAG to clone into pSLIK 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4093)
  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 4093)
  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_p53DD
FEATURES             Location/Qualifiers
     source          1..4093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    42..60
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    78..96
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    114..132
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    149..167
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    185..203
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     250..274
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(628..727)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(839..858)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(917..936)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1510..2043)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2232..2237,2238..2254,2255..2260))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      2917..3505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3406..3425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3670..3756
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3848..3875
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3883..3906
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    3925..4024
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4056..4093,1..277)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4064..4082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gtttactccc tatcagtgat agagaacgat gtcgagttta ctccctatca gtgatagaga
       61 acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
      121 cagtgataga gaacgtatgt cgagtttatc cctatcagtg atagagaacg tatgtcgagt
      181 ttactcccta tcagtgatag agaacgtatg tcgaggtagg cgtgtacggt gggaggccta
      241 tataagcaga gctcgtttag tgaaccgtca gatcgccact agtatgactg ccatggagga
      301 gtcacagtcg gatatcagcc tcgagagagc gctgcccacc tgcacaagcg cctctccccc
      361 gcaaaagaaa aaaccacttg atggagagta tttcaccctc aagatccgcg ggcgtaaacg
      421 cttcgagatg ttccgggagc tgaatgaggc cttagagtta aaggatgccc atgctacaga
      481 ggagtctgga gacagcaggg ctcactccag ctacctgaag accaagaagg gccagtctac
      541 ttcccgccat aaaaaaacaa tggtcaagaa agtggggcct gactcagact gacaattgga
      601 agactaatgc gttttcgaga tatctagacc cagctttctt gtacaaagtt ggcattataa
      661 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
      721 ttatttgcca tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca
      781 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
      841 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
      901 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
      961 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1021 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1081 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1141 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1201 attgttgatg cgctggcagt gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1261 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     1321 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1381 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1441 tcacttgata accttatttt tgacgagggg aaattaggcc gggaagccga tctcggcttg
     1501 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
     1561 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
     1621 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
     1681 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
     1741 ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     1801 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     1861 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     1921 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     1981 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     2041 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     2101 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     2161 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     2221 acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt gccttcatcc
     2281 gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag gcatttctgt
     2341 cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca ttggcggcct
     2401 tgctgttctt ctacggcaag gtgctgtgca cggatctgcc ctggcttcag gagatcggaa
     2461 gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc
     2521 tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat agttctagtg
     2581 gttggctact aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2641 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2701 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2761 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     2821 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3061 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3541 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3601 ccgtattacc gctagcatgg atctcgggga cgtctaacta ctaagcgaga gtagggaact
     3661 gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg ttttatctgt
     3721 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     3781 gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     3841 actaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctgtt
     3901 agttagttac ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
     3961 tcgttgcaac aaattgataa gcaatgcttt tttataatgc caactttgta caaaaaagca
     4021 ggctttaaag gaaccaattc agtcgactgg atcccgagtt tactccctat cagtgataga
     4081 gaacgtatgt cga
//
