LOCUS       sequence-431528-        3601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (C' terminus) p65; HSF1; NpNLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3601)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 3601)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA200
FEATURES             Location/Qualifiers
     source          1..3601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             444..986
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             1011..1382
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             1392..1439
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(1511..1527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1511..1527)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1524..1546)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1535..1551
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1559..1565,1566..1583,1584..1589))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1604..1625
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1742..1759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1913..2501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1993..2012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2679..3488)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3402..3421
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3480..3499
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacggg tccccttcag ggcagatcag caaccaggcc ctggctctgg
      481 cccctagctc cgctccagtg ctggcccaga ctatggtgcc ctctagtgct atggtgcctc
      541 tggcccagcc acctgctcca gcccctgtgc tgaccccagg accaccccag tcactgagcg
      601 ctccagtgcc caagtctaca caggccggcg aggggactct gagtgaagct ctgctgcacc
      661 tgcagttcga cgctgatgag gacctgggag ctctgctggg gaacagcacc gatcccggag
      721 tgttcacaga tctggcctcc gtggacaact ctgagtttca gcagctgctg aatcagggcg
      781 tgtccatgtc tcatagtaca gccgaaccaa tgctgatgga gtaccccgaa gccattaccc
      841 ggctggtgac cggcagccag cggccccccg accccgctcc aactcccctg ggaaccagcg
      901 gcctgcctaa tgggctgtcc ggagatgaag atttctcaag catcgctgat atggacttta
      961 gtgccctgct gtcacagatt tcctctagtg ggcagggagg aggtggaagc ggcttcagcg
     1021 tggacaccag tgccctgctg gacctgttca gcccctcggt gaccgtgccc gacatgagcc
     1081 tgcctgacct tgacagcagc ctggccagta tccaagagct cctgtctccc caggagcccc
     1141 ccaggcctcc cgaggcagag aacagcagcc cggattcagg gaagcagctg gtgcactaca
     1201 cagcgcagcc gctgttcctg ctggaccccg gctccgtgga caccgggagc aacgacctgc
     1261 cggtgctgtt tgagctggga gagggctcct acttctccga aggggacggc ttcgccgagg
     1321 accccaccat ctccctgctg acaggctcgg agcctcccaa agccaaggac cccactgtct
     1381 ccggcaccag caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     1441 gaagcacgtc ttcatcggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgg
     1501 tgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1561 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1621 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1681 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1741 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaagcc
     2641 caatctgaat aatgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
     2701 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     2761 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     2821 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     2881 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2941 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     3001 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     3061 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     3121 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     3181 gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3241 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3301 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3361 ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3421 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata
     3481 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     3541 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc
     3601 a
//
