LOCUS       sequence-272432-        6568 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 6568)
  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 6568)
  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_BirA_NRF2
FEATURES             Location/Qualifiers
     source          1..6568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..315
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    80..98
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    116..134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    152..170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    187..205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    223..241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     288..312
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(322..672,673..675,676..1284)
                     /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"
     protein_bind    complement(3141..3240)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3352..3371)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3430..3449)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(4023..4556)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4745..4750,4751..4767,4768..4773))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      5430..6018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5919..5938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      6183..6269
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6361..6388
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     6396..6419
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    6438..6537
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
       61 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
      121 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
      181 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      241 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
      301 aaccgtcaga tcgccactag aatgaaggac aacaccgtgc ccctgaagct gatcgccctg
      361 ctggccaacg gcgagttcca ctctggcgag cagctgggag agaccctggg aatgagcaga
      421 gccgccatca acaagcacat ccagacactg agagactggg gagtggacgt gttcaccgtg
      481 cctggcaagg gctacagcct gcctgagcct atccagctgc tgaacgccaa gcagatcctg
      541 ggacagctgg atggcggaag cgtggccgtg ctgcctgtga tcgactccac caatcagtac
      601 ctgctggaca gaatcggaga gctgaagtcc ggcgacgcct gcatcgccga gtaccagcag
      661 gctggcagag gaggcagagg acggaagtgg ttcagcccat tcggagccaa cctgtacctg
      721 tccatgttct ggagactgga gcagggacct gctgctgcca tcggactgag tctggtgatc
      781 ggaatcgtga tggccgaggt gctgagaaag ctgggagccg acaaggtgag agtgaagtgg
      841 cctaatgacc tgtacctcca ggaccgcaag ctggctggca tcctggtgga gctgacaggc
      901 aagacaggcg atgccgctca gatcgtgatc ggagccggaa tcaacatggc catgagaaga
      961 gtggaggaga gcgtggtgaa ccagggctgg atcaccctgc aggaggctgg catcaacctg
     1021 gaccggaaca ccctggccgc catgctgatc agagagctga gagccgctct ggagctgttc
     1081 gagcaggagg gactggctcc ttacctgagc agatgggaga agctggacaa cttcatcaac
     1141 agacctgtga agctgatcat cggcgacaag gaaatcttcg gcatctccag aggaatcgac
     1201 aagcagggag ctctgctgct ggagcaggac ggaatcatca agccctggat gggcggagaa
     1261 atctccctga gaagcgcaga gaagactagt atggacttgg agctgccgcc gccgggactc
     1321 ccgtcccagc aggacatgga tttgattgac atactttgga ggcaagatat agatcttgga
     1381 gtaagtcgag aagtatttga cttcagtcag cgacggaaag agtatgagct ggaaaaacag
     1441 aaaaaacttg aaaaggaaag acaagaacaa ctccaaaagg agcaagagaa agcctttttc
     1501 gctcagttac aactagatga agagacaggt gaatttctcc caattcagcc agcccagcac
     1561 atccagtcag aaaccagtgg atctgccaac tactcccagg ttgcccacat tcccaaatca
     1621 gatgctttgt actttgatga ctgcatgcag cttttggcgc agacattccc gtttgtagat
     1681 gacaatgagg tttcttcggc tacgtttcag tcacttgttc ctgatattcc cggtcacatc
     1741 gagagcccag tcttcattgc tactaatcag gctcagtcac ctgaaacttc tgttgctcag
     1801 gtagcccctg ttgatttaga cggtatgcaa caggacattg agcaagtttg ggaggagcta
     1861 ttatccattc ctgagttaca gtgtcttaat attgaaaatg acaagctggt tgagactacc
     1921 atggttccaa gtccagaagc caaactgaca gaagttgaca attatcattt ttactcatct
     1981 ataccctcaa tggaaaaaga agtaggtaac tgtagtccac attttcttaa tgcttttgag
     2041 gattccttca gcagcatcct ctccacagaa gaccccaacc agttgacagt gaactcatta
     2101 aattcagatg ccacagtcaa cacagatttt ggtgatgaat tttattctgc tttcatagct
     2161 gagcccagta tcagcaacag catgccctca cctgctactt taagccattc actctctgaa
     2221 cttctaaatg ggcccattga tgtttctgat ctatcacttt gcaaagcttt caaccaaaac
     2281 caccctgaaa gcacagcaga attcaatgat tctgactccg gcatttcact aaacacaagt
     2341 cccagtgtgg catcaccaga acactcagtg gaatcttcca gctatggaga cacactactt
     2401 ggcctcagtg attctgaagt ggaagagcta gatagtgccc ctggaagtgt caaacagaat
     2461 ggtcctaaaa caccagtaca ttcttctggg gatatggtac aacccttgtc accatctcag
     2521 gggcagagca ctcacgtgca tgatgcccaa tgtgagaaca caccagagaa agaattgcct
     2581 gtaagtcctg gtcatcggaa aaccccattc acaaaagaca aacattcaag ccgcttggag
     2641 gctcatctca caagagatga acttagggca aaagctctcc atatcccatt ccctgtagaa
     2701 aaaatcatta acctccctgt tgttgacttc aacgaaatga tgtccaaaga gcagttcaat
     2761 gaagctcaac ttgcattaat tcgggatata cgtaggaggg gtaagaataa agtggctgct
     2821 cagaattgca gaaaaagaaa actggaaaat atagtagaac tagagcaaga tttagatcat
     2881 ttgaaagatg aaaaagaaaa attgctcaaa gaaaaaggag aaaatgacaa aagccttcac
     2941 ctactgaaaa aacaactcag caccttatat ctcgaagttt tcagcatgct acgtgatgaa
     3001 gatggaaaac cttattctcc tagtgaatac tccctgcagc aaacaagaga tggcaatgtt
     3061 ttccttgttc ccaaaagtaa gaagccagat gttaagaaaa actagcaatt ggaagactaa
     3121 tgcgttttcg agatatctag acccagcttt cttgtacaaa gttggcatta taagaaagca
     3181 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3241 ccatccagct gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     3301 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     3361 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     3421 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3481 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3541 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3601 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3661 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3721 atgcgctggc agtgtccctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3781 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3841 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3901 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3961 ataaccttat ttttgacgag gggaaattag gccgggaagc cgatctcggc ttgaacgaat
     4021 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     4081 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     4141 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     4201 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     4261 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     4321 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     4381 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     4441 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     4501 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     4561 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     4621 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     4681 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     4741 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     4801 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gaggcatttc tgtcctggct
     4861 ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag gcattggcgg ccttgctgtt
     4921 cttctacggc aaggtgctgt gcacggatct gccctggctt caggagatcg gaagacctcg
     4981 gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa gtggttcgca tcctcggttt
     5041 tctggaaggc gagcatcgtt tgttcgccca ggactctagc tatagttcta gtggttggct
     5101 actaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     5161 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     5221 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     5281 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     5341 ggacggcgca agctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6121 accgctagca tggatctcgg ggacgtctaa ctactaagcg agagtaggga actgccaggc
     6181 atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt
     6241 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc
     6301 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc
     6361 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttcct gttagttagt
     6421 tacttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
     6481 aacaaattga taagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggcttta
     6541 aaggaaccaa ttcagtcgac tggatccc
//
