LOCUS       sequence-420418-        4124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homologous donor template for NFKB1 knockout expressing EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4124)
  AUTHORS   Sato Y, Osada E, Manome Y
  TITLE     Non-canonical NFKB signaling endows suppressive function through 
            FOXP3-dependent regulatory T cell program.
  JOURNAL   Heliyon. 2023 Nov 26;9(12):e22911. doi: 
            10.1016/j.heliyon.2023.e22911. eCollection 2023 Dec.
  PUBMED    38125410
REFERENCE   2  (bases 1 to 4124)
  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: "Heliyon."; 
            date: "2023-11-26"; volume: "9(12):e22911. doi"; pages: " 
            10.1016/j.heliyon.2023.e22911. eCollection 2023 Dec"
COMMENT     Sequence Label: pUCmini-NFKB1-PGK-EGFP
FEATURES             Location/Qualifiers
     source          1..4124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        473..983
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     902..920
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      999..1008
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1005..1007,1008..1010,1011..1724)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1005..1007,1008..1010,1011..1721)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1050..1071)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1311..1330)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1658..1679
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1731..1842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2369..2957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2449..2468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3128..3919,3920..3988))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3751..3770
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3989..4093)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgatgc ggccgccctg ccttcttcct ttgatttcca tcaaccacca gtttctttct
       61 catcaatcat ttacccagct tttgtgacca agaattttga tgaggatgca gaggaaggca
      121 tgttcttggt taataggaaa ggaggcttca tgacaacaca gcatcctcta agatgtgttg
      181 ctccaacttt cacaatacat attgtgtttt aagtggcctg cttcttggag tgtatcttcc
      241 cactacacaa aacaaccacc attaacattt tcatttactt ccaccctgtc atttgttcat
      301 cacatacttt taaaaacagg gctgtaatca tgtatataca tatatgaata actttatatt
      361 gtacttttta tacttactgg cttatcataa acatgtttcc atgttgctgg agagtcagat
      421 atgctaactt ttggaatgtg cttcttatat agatctgcaa ctgaattccc acggggttgg
      481 ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg
      541 gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca
      601 gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc
      661 ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg
      721 cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac
      781 cgcgatgggc tgtggccaat agcggctgct cagcggggcg cgccgagagc agcggccggg
      841 aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc
      901 ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc
      961 gaatcaccga cctctctccc cagggggatc caccggtcgc caccatggtg agcaagggcg
     1021 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1081 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1141 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1201 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     1261 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1321 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1381 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1441 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     1501 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     1561 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     1621 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1681 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagtcgac ctgtgccttc
     1741 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1801 cactcccact gtcctttcct aataaaatga ggaaattgca tcatgtcgga ccagcaaagg
     1861 ttattgttca gttggtcaca aatggaaaaa atatccacct gcatgcccac agcctggtgg
     1921 gaaaacactg tgaggatggg atctgcactg taactgctgg acccaaggac atggtggtcg
     1981 ggtaagtagg ggtatatgat gctgtggaag gtaggaacag atagaatatg ggatgcagga
     2041 acttagaatg aacaggcccc tttcattagg gacctttctg agaccctcag atgacctcaa
     2101 aaaactgtgt aaacttgtgc tttcatttgg actccaggaa gtaaggtgac cacatgtctg
     2161 gatttgcttg agacagcccg gtttgcccct gacctaattg tttatagcac cctcttctac
     2221 tcttaaaggt ggcccaattt ggatgatcac ttatatgtgg tcacttgacc tacaaggtat
     2281 ttgtgccacg cggccgcatc ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2341 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2401 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2461 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2521 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2581 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2641 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2701 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2761 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2821 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2881 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2941 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3001 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3061 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3121 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3181 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3241 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3301 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3361 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3421 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3481 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3541 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3601 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3661 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3721 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3781 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3841 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3901 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3961 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4021 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4081 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
