LOCUS       sequence-420419-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homologous donor template for NFKB1 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-NFKB1-PGK-tdTomato
FEATURES             Location/Qualifiers
     source          1..4835
                     /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             1005..2435
                     /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    2442..2553
                     /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 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 aggaggtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc
     1081 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     1141 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
     1201 tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc
     1261 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg
     1321 tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc
     1381 gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1441 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc
     1501 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga
     1561 agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca
     1621 acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc
     1681 tggggcatgg caccggcagc accggcagcg gcagctccgg caccgcctcc tccgaggaca
     1741 acaacatggc cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga
     1801 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1861 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     1921 cccagttcat gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca
     1981 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2041 gtctggtgac cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga
     2101 agatgcgcgg caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct
     2161 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc
     2221 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg
     2281 ccaagaagcc cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct
     2341 cccacaacga ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc
     2401 tgttcctgta cggcatggac gagctgtaca agtaggtcga cctgtgcctt ctagttgcca
     2461 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2521 tgtcctttcc taataaaatg aggaaattgc atcatgtcgg accagcaaag gttattgttc
     2581 agttggtcac aaatggaaaa aatatccacc tgcatgccca cagcctggtg ggaaaacact
     2641 gtgaggatgg gatctgcact gtaactgctg gacccaagga catggtggtc gggtaagtag
     2701 gggtatatga tgctgtggaa ggtaggaaca gatagaatat gggatgcagg aacttagaat
     2761 gaacaggccc ctttcattag ggacctttct gagaccctca gatgacctca aaaaactgtg
     2821 taaacttgtg ctttcatttg gactccagga agtaaggtga ccacatgtct ggatttgctt
     2881 gagacagccc ggtttgcccc tgacctaatt gtttatagca ccctcttcta ctcttaaagg
     2941 tggcccaatt tggatgatca cttatatgtg gtcacttgac ctacaaggta tttgtgccac
     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
//
