LOCUS       sequence-431448-        3816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (N' terminus) activator from p65 subunit of mouse 
            Nfkb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3816)
  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 3816)
  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: AA125
FEATURES             Location/Qualifiers
     source          1..3816
                     /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"
     regulatory      444..453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             456..998
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             1635..1655
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1726..1742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1726..1742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1739..1761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1750..1766
                     /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(1774..1780,1781..1798,1799..1804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1819..1840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1957..1974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2128..2716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2208..2227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2894..3703)
                     /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     3617..3636
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3695..3714
                     /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 tgaagacgta ggagccacca tggccccttc tggacagatc agtaatcagg
      481 ccctggccct ggcccctagc tctgccccag tgctggccca aacaatggtc cctagcagcg
      541 ccatggtgcc cctggctcaa cctcctgccc cagcccctgt gctgacacct ggacctcctc
      601 agagcctgtc cgcccctgtg ccaaaaagca cacaggctgg cgaaggcacc ctgagcgagg
      661 ccctgctgca cctgcagttc gacgctgacg aggatctcgg cgccctgctg ggcaacagca
      721 ccgatcccgg cgttttcacc gacctggctt ctgtggacaa cagcgaattc cagcagctgc
      781 tgaaccaggg cgtgtccatg agccacagca cagccgaacc catgctgatg gaataccccg
      841 aagccatcac ccggctggtc accggatctc agagaccccc cgacccagcc cctacacctc
      901 tgggaaccag cggcctgcct aacggcctga gcggagatga agatttcagc agcattgccg
      961 acatggactt cagcgctctg ctgagccaga tctcctctag cggccaagga ggcggcggct
     1021 ctggttttag cgtggacacc agcgcacttc tggacctgtt ttctcctagc gtgaccgtgc
     1081 ctgatatgag cctgcctgat ctggatagca gcctcgcctc tatccaggag ctgctgtctc
     1141 cccaggagcc ccctagacct ccagaggccg agaatagcag ccctgacagc ggcaagcagc
     1201 tggtgcacta caccgcccag cccctgttcc tgctggaccc cggctccgtg gataccggct
     1261 ccaacgacct gcctgtgctg ttcgagctgg gcgagggctc ctacttcagc gaaggcgacg
     1321 gcttcgccga ggaccctacc atcagcctgc tgacaggcag cgagcctcct aaggccaagg
     1381 accccaccgt gtccggaggg ccgagctctg gcgcaccccc accaagtgga gggtctcctg
     1441 ccgggtcccc aacatctact gaagaaggca ccagcgaatc cgcaacgccc gagtcaggcc
     1501 ctggtacctc cacagaacca tctgaaggta gtgcgcctgg ttccccagct ggaagcccta
     1561 cttccaccga agaaggcacg tcaaccgaac caagtgaagg atctgcccct gggaccagca
     1621 ctgaaccatc tgagccaaaa aagaagagaa aggtaggcag cagtcttcat cggatcccgg
     1681 gcccgtcgac tgcagaggcc tgcatgcaag cttggtgtaa tcatggtcat agctgtttcc
     1741 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     1801 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1861 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1921 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1981 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2041 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2101 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2161 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2221 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2281 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2341 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2401 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2461 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2521 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2581 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2641 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2761 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2821 ccttttaaat taaaaatgaa gttttaaatc aagcccaatc tgaataatgt tacaaccaat
     2881 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     2941 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     3001 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     3061 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     3121 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     3181 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     3241 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     3301 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     3361 ctgaatcagg atattcttct aatacctgga atgctgtttt tccggggatc gcagtggtga
     3421 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     3481 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     3541 catgtttcag aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac
     3601 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     3661 aatttaatcg cggcctcgac gtttcccgtt gaatatggct cataacaccc cttgtattac
     3721 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt
     3781 aacatcagag attttgagac acgggccaga gctgca
//
