LOCUS       sequence-272436-        4717 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 4717)
  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 4717)
  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
FEATURES             Location/Qualifiers
     source          1..4717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..401
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    95..113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    131..149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    166..184
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    202..220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    238..256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    273..291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    309..327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     374..398
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(408..758,759..761,762..1370)
                     /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(1376..1475)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1587..1606)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1665..1684)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(2258..2791)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2980..2985,2986..3002,3003..3008))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      3665..4253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4154..4173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4418..4504
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4596..4623
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4631..4654
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    join(4673..4717,1..55)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 caaattgata agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctttaaa
       61 ggaaccaatt cagtcgactg gatcccgagt ttactcccta tcagtgatag agaacgtatg
      121 tcgagtttac tccctatcag tgatagagaa cgatgtcgag tttactccct atcagtgata
      181 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc
      241 ctatcagtga tagagaacgt atgtcgagtt tatccctatc agtgatagag aacgtatgtc
      301 gagtttactc cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggagg
      361 cctatataag cagagctcgt ttagtgaacc gtcagatcgc cactagaatg aaggacaaca
      421 ccgtgcccct gaagctgatc gccctgctgg ccaacggcga gttccactct ggcgagcagc
      481 tgggagagac cctgggaatg agcagagccg ccatcaacaa gcacatccag acactgagag
      541 actggggagt ggacgtgttc accgtgcctg gcaagggcta cagcctgcct gagcctatcc
      601 agctgctgaa cgccaagcag atcctgggac agctggatgg cggaagcgtg gccgtgctgc
      661 ctgtgatcga ctccaccaat cagtacctgc tggacagaat cggagagctg aagtccggcg
      721 acgcctgcat cgccgagtac cagcaggctg gcagaggagg cagaggacgg aagtggttca
      781 gcccattcgg agccaacctg tacctgtcca tgttctggag actggagcag ggacctgctg
      841 ctgccatcgg actgagtctg gtgatcggaa tcgtgatggc cgaggtgctg agaaagctgg
      901 gagccgacaa ggtgagagtg aagtggccta atgacctgta cctccaggac cgcaagctgg
      961 ctggcatcct ggtggagctg acaggcaaga caggcgatgc cgctcagatc gtgatcggag
     1021 ccggaatcaa catggccatg agaagagtgg aggagagcgt ggtgaaccag ggctggatca
     1081 ccctgcagga ggctggcatc aacctggacc ggaacaccct ggccgccatg ctgatcagag
     1141 agctgagagc cgctctggag ctgttcgagc aggagggact ggctccttac ctgagcagat
     1201 gggagaagct ggacaacttc atcaacagac ctgtgaagct gatcatcggc gacaaggaaa
     1261 tcttcggcat ctccagagga atcgacaagc agggagctct gctgctggag caggacggaa
     1321 tcatcaagcc ctggatgggc ggagaaatct ccctgagaag cgcagagaag actagaccca
     1381 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     1441 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagc tctggcccgt
     1501 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1561 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1621 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1681 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1741 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1801 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1861 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     1921 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt ccctgcgccg
     1981 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2041 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2101 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2161 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2221 attaggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
     2281 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
     2341 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
     2401 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga
     2461 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
     2521 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
     2581 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
     2641 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
     2701 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
     2761 gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
     2821 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag
     2881 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
     2941 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct
     3001 tatgtcaact gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
     3061 ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc
     3121 tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg
     3181 gatctgccct ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccggtggtg
     3241 ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc
     3301 gcccaggact ctagctatag ttctagtggt tggctactaa taggttgtat tgatgttgga
     3361 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3421 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3481 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3541 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3601 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3661 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3721 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3781 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3841 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3901 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3961 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4021 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4081 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4141 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4201 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4261 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4321 ctgcgttatc ccctgattct gtggataacc gtattaccgc tagcatggat ctcggggacg
     4381 tctaactact aagcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     4441 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     4501 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
     4561 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
     4621 ttttgcgttt ctacaaactc ttcctgttag ttagttactt aagctcgggc cccaaataat
     4681 gattttattt tgactgatag tgacctgttc gttgcaa
//
