LOCUS       sequence-445719-        4502 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Expresses estrogen receptor linked KLF2 and mCherry under EF1a 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4502)
  AUTHORS   Fagerberg E, Attanasio J, Dien C, Singh J, Kessler EA, Abdullah L, 
            Shen J, Hunt BG, Connolly KA, De Brouwer E, He J, Iyer NR, Buck J, 
            Borr ER, Damo M, Foster GG, Giles JR, Huang YH, Tsang JS, 
            Krishnaswamy S, Cui W, Joshi NS
  TITLE     KLF2 maintains lineage fidelity and suppresses CD8 T cell exhaustion
            during acute LCMV infection.
  JOURNAL   Science. 2025 Jan 2;387(6735):eadn2337. doi: 
            10.1126/science.adn2337. Epub 2025 Feb 14.
  PUBMED    39946463
REFERENCE   2  (bases 1 to 4502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2025-01-2"; volume: "387(6735):eadn2337. doi"; pages: " 
            10.1126/science.adn2337. Epub 2025 Feb 14"
COMMENT     Sequence Label: EF1a-ER-KLF2-T2A-mCHERRY
FEATURES             Location/Qualifiers
     source          1..4502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    295..636
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             703..1119
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1049..1071
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1087..1103
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1182..1393
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1407..1416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1416..2351
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             3429..3482
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3489..4193
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWDASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3631..3649)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3899..3918
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4230..4249)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
       61 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
      121 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggact cgctgatcct
      181 tgggagggtc tcctcagatt gattgactgc ccacctcggg ggtctttcat ttggaggttc
      241 caccgagatt tggagacccc tgcccaggga ccaccgaccc ccccgccggg aggtaagctg
      301 gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg catctaatgt
      361 ttgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      421 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
      481 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
      541 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
      601 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
      661 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
      721 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
      781 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
      841 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
      901 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
      961 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
     1021 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
     1081 tcctcccttt atccagccct cactccttct ctaggcgccg gaattgcttc gcgatgtacg
     1141 ggccaggaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     1201 cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg
     1261 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     1321 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     1381 cgccagaaca caggaccggt tctagagcca ccatggctgg agacatgaga gctgccaacc
     1441 tttggccaag cccgctcatg atcaaacgct ctaagaagaa cagcctggcc ttgtccctga
     1501 cggccgacca gatggtcagt gccttgttgg atgctgagcc ccccatactc tattccgagt
     1561 atgatcctac cagacccttc agtgaagctt cgatgatggg cttactgacc aacctggcag
     1621 acagggagct ggttcacatg atcaactggg cgaagagggt gccaggcttt gtggatttga
     1681 ccctccatga tcaggtccac cttctagaat gtgcctggct agagatcctg atgattggtc
     1741 tcgtctggcg ctccatggag cacccagtga agctactgtt tgctcctaac ttgctcttgg
     1801 acaggaacca gggaaaatgt gtagagggca tggtggagat cttcgacatg ctgctggcta
     1861 catcatctcg gttccgcatg atgaatctgc agggagagga gtttgtgtgc ctcaaatcta
     1921 ttattttgct taattctgga gtgtacacat ttctgtccag caccctgaag tctctggaag
     1981 agaaggacca tatccaccga gtcctggaca agatcacaga cactttgatc cacctgatgg
     2041 ccaaggcagg cctgaccctg cagcagcagc accagcggct ggcccagctc ctcctcatcc
     2101 tctcccacat caggcacatg agtaacaaag gcatggagca tctgtacagc atgaagtgca
     2161 agaacgtggt gcccctctat gacctgctgc tggaggcggc ggacgcccac cgcctacatg
     2221 cgcccactag ccgtggaggg gcatccgtgg aggaaacgga ccaaagccac ttggccactg
     2281 cgggctctac ttcatcgcat tccttgcaaa agtattacat cacgggggag gcagagggtt
     2341 tccctgccac agctgtcgac atggcgctca gcgagcctat cttgcctagt tttgccactt
     2401 ttgcatcccc ctgtgaacga ggattgcagg aacgctggcc tcgcaacgaa ccagaagctg
     2461 gagggaccga cgaagacctc aacaatgtgt tggattttat cctgagtatg gggctcgatg
     2521 gcttgggggc cgaaaaccca ccagaaccac ctccacagcc ccctccacca gccttttatt
     2581 acccagaacc cggagcacca ccaccatact caatccccgc cgcttcactt ggtacagaac
     2641 tcctcaggcc agatttggat cctccacagg gtcccgcatt gcacggacgc ttccttttgg
     2701 cccctccagg gcgactggtc aaggccgagc ctcctgaagt ggatggtgga ggctatgggt
     2761 gcgctccagg gctggctcac gggccaagag gcttgaaact tgagggcgct ccaggtgcaa
     2821 ccggagcctg catgcggggt cccgccggcc gccccccgcc gcccccggac acgccgccgc
     2881 tcagccccga cggccccctg cgcatcccgg cgtccggtcc acgcaacccg ttcccgccgc
     2941 ccttcggtcc cggccccagc ttcggcggcc ccggccccgc gttgcactac gggcctcccg
     3001 cgcctggcgc cttcggtctt ttcgaggacg cggcggcagc ggcggcggcg ctgggcttgg
     3061 ctccacctgc cacgcgcggt ctcctcacgc cgccctcgtc cccgctggag ctgctggagg
     3121 ccaagcccaa acgcggccgc cgctcctggc cccgcaagcg cgccgccaca catacttgca
     3181 gctacaccaa ctgcggcaag acctacacca agagctcgca cctaaaggcg catctgcgta
     3241 cacacacagg tgagaagcct tatcattgca actgggaagg atgcggctgg aagttcgcgc
     3301 gctctgacga gcttacccgc cactaccgaa agcacacagg tcacagaccc tttcagtgcc
     3361 acttgtgcga ccgggccttc tccaggtccg accacctggc cttgcacatg aagcgacaca
     3421 tggctagcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     3481 ctggtaccgt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     3541 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     3601 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     3661 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     3721 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     3781 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     3841 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     3901 ccgtaatgca gaagaagacc atgggctggg atgcctcctc cgagcggatg taccccgagg
     3961 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     4021 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     4081 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     4141 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagaccggtt
     4201 agtaatgagt ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaaggg
     4261 gccgcgactc tagagtcgac ctgcaggcat gcaagcttca ggtagccggc taacgttaac
     4321 aaccggtacc tctagaacta tagctagcat gcgcaaattt aaagcgctga tatcgataaa
     4381 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     4441 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata
     4501 ga
//
