LOCUS       sequence-431540-        3205 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Fragmid fragment: (C' terminus) p65; NpNLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3205)
  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 3205)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: AA199
FEATURES             Location/Qualifiers
     source          1..3205
                     /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             996..1043
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(1115..1131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1115..1131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1128..1150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1139..1155
                     /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(1163..1169,1170..1187,1188..1193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1208..1229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1346..1363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1517..2105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1597..1616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2283..3092)
                     /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     3006..3025
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3084..3103
                     /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 ggagatgagg acttctcaag catcgctgat atggacttta
      961 gtgccctgct gtcacagatt tcctctggca ccagcaaaag gccggcggcc acgaaaaagg
     1021 ccggccaggc aaaaaagaaa aagggaagca cgtcttcatc ggatcccggg cccgtcgact
     1081 gcagaggcct gcatgcaagc ttggtgtaat catggtcata gctgtttcct gtgtgaaatt
     1141 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1201 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1261 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1321 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1381 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1441 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2221 aaaaatgaag ttttaaatca agcccaatct gaataatgtt acaaccaatt aaccaattct
     2281 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     2341 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2401 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2461 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2521 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     2581 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     2641 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     2701 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     2761 tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag taaccatgca
     2821 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     2881 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     2941 aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
     3001 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3061 ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     3121 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     3181 ttttgagaca cgggccagag ctgca
//
