LOCUS       sequence-420420-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homologous donor template for NFKB2 knockout expressing tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4835)
  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 4835)
  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-tdTomato
FEATURES             Location/Qualifiers
     source          1..4835
                     /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             1013..2443
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    2450..2561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3080..3668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3160..3179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3839..4630,4631..4699))
                     /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     4462..4481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4700..4804)
                     /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 gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg agggctccat
     1081 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1141 gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc
     1201 cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca tccccgatta
     1261 caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1321 cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga tctacaaggt
     1381 gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg
     1441 ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca
     1501 ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga ccatctacat
     1561 ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac
     1621 ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg gccgccacca
     1681 cctgttcctg gggcatggca ccggcagcac cggcagcggc agctccggca ccgcctcctc
     1741 cgaggacaac aacatggccg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg
     1801 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1861 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     1921 cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc
     1981 cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2041 ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta
     2101 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
     2161 catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
     2221 gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat
     2281 ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga
     2341 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
     2401 ccaccacctg ttcctgtacg gcatggacga gctgtacaag taggtcgacc tgtgccttct
     2461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2521 actcccactg tcctttccta ataaaatgag gaaattgcat ctctgtaact acgagggacc
     2581 agccaagatc gaggtggacc tggtaacaca cagtgaccca cctcgtgctc atgcccacag
     2641 tctggtgggc aagcaatgct cggagctggg gatctgcgcc gtttctgtgg ggcccaagga
     2701 catgactgcc cagtaggtgc cctctacgcc tggcccccac tggtatgccc gtctgccagt
     2761 cccaggcccc agcccacctc catgagctta gcatctgacc aaggggaaag atgtaggttg
     2821 gccccaaacc caagggccta agtagaaact ccaatggctt ccttgaggaa gtaaggctga
     2881 gctgagccct ggcaatggga aaggtgcctc aggaagaaag aactgcatgg ccaaaggcct
     2941 ccgattctct cttctcagat ttaacaacct gggtgtcctg catgtgacta agaagaacat
     3001 gcggccgcat cggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4801 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
