LOCUS       sequence-420417-        4124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homologous donor template for NFKB2 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-NFKB2-PGK-EGFP
FEATURES             Location/Qualifiers
     source          1..4124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        481..991
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     910..928
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      1007..1016
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1013..1015,1016..1018,1019..1732)
                     /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(1013..1015,1016..1018,1019..1729)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1058..1079)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1319..1338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1666..1687
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1739..1850
                     /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 gtgatcgtgg aacagcctaa gcaggtgagt gagcaaaagg
       61 gagggtgtgg aatggcttca gctttgggga caaatggggt agtggtagct ggctggccat
      121 ggaggagcca ttgccgaagg aggccacagg ggattggatg gtcactgctg ctgatcagag
      181 tgctgtagtt tggttcaggg ctactaccag gcactgcggt cactgctggc ctgggtggtc
      241 ttccctgatc acaatgctac tatgcccttg gaccttcaga gaggcttccg atttcgatat
      301 ggctgtgaag gcccctccca tggaggactg cccggtgcct ccagtgagaa gggccgaaag
      361 acctatccca ctgtcaaggt gagccaggat ggtgctggag ggtgggctaa gtggacagca
      421 tgcccaaggc cctgactgac agtccctgcc tctcgtagat gcggtgggct gaattcccac
      481 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
      541 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
      601 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
      661 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
      721 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
      781 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag
      841 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
      901 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
      961 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggtcgcca ccatggtgag
     1021 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1081 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1141 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1201 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1261 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1321 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1381 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1441 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1501 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1561 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1621 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1681 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagtcgacct
     1741 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1801 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc tctgtaacta
     1861 cgagggacca gccaagatcg aggtggacct ggtaacacac agtgacccac ctcgtgctca
     1921 tgcccacagt ctggtgggca agcaatgctc ggagctgggg atctgcgccg tttctgtggg
     1981 gcccaaggac atgactgccc agtaggtgcc ctctacgcct ggcccccact ggtatgcccg
     2041 tctgccagtc ccaggcccca gcccacctcc atgagcttag catctgacca aggggaaaga
     2101 tgtaggttgg ccccaaaccc aagggcctaa gtagaaactc caatggcttc cttgaggaag
     2161 taaggctgag ctgagccctg gcaatgggaa aggtgcctca ggaagaaaga actgcatggc
     2221 caaaggcctc cgattctctc ttctcagatt taacaacctg ggtgtcctgc atgtgactaa
     2281 gaagaacatg 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
//
