LOCUS       sequence-252963-        9947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knock-in of TagBFP into mouse Esrrb locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9947)
  AUTHORS   Benchetrit H, Jaber M, Zayat V, Sebban S, Pushett A, Makedonski K, 
            Zakheim Z, Radwan A, Maoz N, Lasry R, Renous N, Inbar M, Ram O, 
            Kaplan T, Buganim Y
  TITLE     Direct Induction of the Three Pre-implantation Blastocyst Cell Types
            from Fibroblasts.
  JOURNAL   Cell Stem Cell. 2019 Jun 6;24(6):983-994.e7. doi: 
            10.1016/j.stem.2019.03.018. Epub 2019 Apr 25.
  PUBMED    31031139
REFERENCE   2  (bases 1 to 9947)
  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"; doi: 
            "10.1016/j.stem.2019.03.018"; journalName: "Cell Stem Cell"; date: 
            "2019-06-6- 6"; volume: "24"; issue: "6"; pages: "983-994.e7"
COMMENT     Sequence Label: pNTKV- Esrrb-KI-3'UTR-2A-TagBFP-RBGpA
FEATURES             Location/Qualifiers
     source          1..9947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        116..178
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             257..1387
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMIQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(1385..1404)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1429..1477
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1909..1925
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4972..5025
                     /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             5041..5742
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     intron          5771..6343
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      8008..8017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        8061..8165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8166..8234,8235..9026)
                     /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     complement(8384..8403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9197..9785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9686..9705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctccacccag gcctggaatg tttccaccca atgtcgagca aaccccgccc agcgtcttgt
       61 cattggcgaa ttggaacacg cagatgcagt cggggcggcg cggtcccagg tccacttcgc
      121 atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaca
      181 gcgtcaacag cgtgccgcac atcttggtgg cgtgaaactc ccgcacctct tcggccagcg
      241 ccttgtagaa gcgcgtatgg cttcgtaccc cggccatcag cacgcgtctg cgttcgacca
      301 ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg ttgcgccctc gccggcagca
      361 agaagccacg gaagtccgcc cggagcagaa aatgcccacg ctactgcggg tttatataga
      421 cggtccccac gggatgggga aaaccaccac cacgcaactg ctggtggccc tgggttcgcg
      481 cgacgatatc gtctacgtac ccgagccgat gacttactgg cgggtgctgg gggcttccga
      541 gacaatcgcg aacatctaca ccacacaaca ccgccttgac cagggtgaga tatcggccgg
      601 ggacgcggcg gtggtaatga caagcgccca gataacaatg ggcatgcctt atgccgtgac
      661 cgacgccgtt ctggctcctc atatcggggg ggaggctggg agctcacatg ccccgccccc
      721 ggccctcacc ctcatcttcg accgccatcc catcgccgcc ctcctgtgct acccggccgc
      781 gcgatacctt atgggcagca tgacccccca ggccgtgctg gcgttcgtgg ccctcatccc
      841 gccgaccttg cccggcacaa acatcgtgtt gggggccctt ccggaggaca gacacatcga
      901 ccgcctggcc aaacgccagc gccccggcga gcggcttgac ctggctatgc tggccgcgat
      961 tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat ctgcagggcg gcgggtcgtg
     1021 gcgggaggat tggggacagc tttcggggac ggccgtgccg ccccagggtg ccgagcccca
     1081 gagcaacgcg ggcccacgac cccatatcgg ggacacgtta tttaccctgt ttcgggcccc
     1141 cgagttgctg gcccccaacg gcgacctgta caacgtgttt gcctgggcct tggacgtctt
     1201 ggccaaacgc ctccgtccca tgcacgtctt tatcctggat tacgaccaat cgcccgccgg
     1261 ctgccgggac gccctgctgc aacttacctc cgggatgatc cagacccacg tcaccacccc
     1321 aggctccata ccgacgatct gcgacctggc gcgcacgttt gcacgggaga tgggggaggc
     1381 taactgaaac acggaaggag acaataccgg aaggaacctg cgctatgacg gcaataaaaa
     1441 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     1501 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     1561 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     1621 ggggcggcag gccctgccat agccacgggc cccgtgggtt agggacgggg tcccccatgg
     1681 ggaatggttt atggttcgtg ggggttatta ttttgggcgt tgcgtggggt cagtccacga
     1741 ctggactgag cagacagacc catggttttt ggatggcctg ggcatggacc gcatgtactg
     1801 gcgcgacacg aacaccgggc gtctgtggct gccaaacacc cccgaccccc aaaaaccacc
     1861 gcgcggattt ctggcgccgc cggaccaact aaacctgact acggaccggt aaaacgacgg
     1921 ccagtgttaa ctgctgggga aagagagaca tacagggtct ctatgtgcat gagagtaaca
     1981 ttagaaacac gcacatacaa ttgtaattcc actgcagggg cactacgtgg cagagttctt
     2041 gctctggaca gggccaggtg gacctgagtt caaatcctgt ctctgaggct gaagagagag
     2101 atggtttagc aattaagagc accggctgct cttgcagagg accagagttt agttcccagc
     2161 actcacaaac ttccaattct aactccaaag gatccaatgc cctcttttgg ccttcactgg
     2221 cactgcatac acatggcata tatacatact ggcacacaca cacacactct gtcacactaa
     2281 cagcatgacc ttggttagct ctgtccttga gtaggaggta ttcccagata cctcaagtac
     2341 ctgtgagctt taaagatcaa tttcggataa tgcagtccag tgcctggcct agaggagcac
     2401 tcagtaaata ttgattattg tccctgctgt tacaatgtca tgagaaatgg tgttcccagc
     2461 acatgatgca ggaaagctga ctcccaggta gagacactcg taccagcaga ctaccgcttc
     2521 actgtttgtg ctgggctttc cccgaaagtc tcttatctca ggaacaccca atccttagca
     2581 acttggcttt gctgctgtta gctccccaga aacaccctct gctgcctttc ctgaggtcag
     2641 gcattgtcag ccgatggaga cttcacacag cccagcagtc tgccttggac atctgtgttg
     2701 gttcaggcct tgacattgta ttgttccttt tgacttcctg cctccttttc cttctgtaat
     2761 cctcttccac tccctccccc tttcccctcc tcctttcccc tccccttccc cctccataaa
     2821 tctttaaccc taaacccctg ccttaggctt tgctcctaag gacctaacct aatacctgtt
     2881 gttgcgtttt cttttttgct gtagctgtgt aaacgaacag cgcatggaat tccatgtgtc
     2941 ctccctgtcc ctccccatcc tctgtacact ctctccactc ttcctgttag gtaatttgct
     3001 gcaattagta agctaatgag gactgacgcc tctccagttt ttgagacagt ttcaatctgt
     3061 cgcccaggct ggcctgggat tgactctata tctcaggctg attcgaaact cacagaaatc
     3121 ctcctgcatc agccttccaa gtattgtaat tataggtgtt taccaaccac tcatggcagt
     3181 ggcccttgcc tgctaccaga gactccttac tcctgccttc cataggtttc taaaacccag
     3241 taccttgtgg tctaggaagg gagatctgga tggacatgcc cctgcaggtc acagtcctga
     3301 ggcagaacag tgacccagga gcctacagag catattggat ggagaaggga tgtctaggag
     3361 accaatcaga cgcccccacc ccccatggct aaagtggata gggtgactca ggtacctggc
     3421 agaagagcaa agctcatcag aggctgaccg ggtccccttg agaatgtgcc agctgtcact
     3481 cacttgcatc ccagccatct ctagggctct ctgacagcag aaaaaaaaaa ttcatagcca
     3541 ttgattttgt aattaacagt ttatcaatgc cattgataca gacttgctga tggattgcaa
     3601 agaaattgat cttgcaatgt tattttattt catttttggt tgaaagcacc tgccaattgt
     3661 ggctccttgg ggagggtgac aggatggaaa atggggtcag aggtggctgg ggcccagccc
     3721 tgtaggaatt ccggaaagta caggctggga ggagatcccc agggggcatg agcaagcatc
     3781 gctatgccca gccagctgat gcagctgcct cggcgctttg gttcaggcga gctctgaact
     3841 tgtgcactcc aatgaacaca gaaggctggc tcattgccct gtgtccacac agcaagcttt
     3901 gaggctccag ttcatgcagg catcacgggt tgttttaaaa aaagtgtttg ggtagttctc
     3961 ccacaagact gggaggttcc ctttgttttg ctcagaccta gctgtatccc aatgtttcag
     4021 acggccggcc gagttcatgg gctgtaagca tcccaagccg atggtgctaa ggatccagat
     4081 tcaagcaagg ccaagcaagg gatacactca tccctgaacg atatccccgc cggctccctc
     4141 tatcagcaca ccctcccctc gcccccaaac acctagacat tgacttccat ccctttatat
     4201 ctgttctgag ttgtaaggtt ggtggctgta ctgcatgtga gtggtaccca tggcccagca
     4261 tcatcatgaa gacgtgatga tgtcagggtt aggtgatagc cgtgaggttc ctaagaccca
     4321 ggtgcttaat tgttgctgta taactgtatg aggtaggtat ggctaggaaa atgggggaca
     4381 gagaagttat ataatgtctc acataactca aaagggactg ccagccagat cccagagtcc
     4441 cccgaacgcc tctgagacat ccaataggtc tgcccatgtt catcggcagt gccctcctgg
     4501 tggtctctct atgagatctg agctcagttc taccctcctc agtctctgac tcagccctgc
     4561 atcaaagatc ctcattctac tctgaatagt cagtcagcag acagacagac agagacaggc
     4621 tggctgctgg tctgtcctcc actccagaaa tgccttccgt gttggggggt ggcagccagc
     4681 ccctgctgac actgcccctc tcctgtgtcc acagattcga tgtacattga gaacctggag
     4741 gcggtgcaga agctccagga cctgctgcac gaggcgctgc aggactatga gctgagtcag
     4801 cgccacgagg agccgcggag ggccggcaag ctgctgctga cgctgcccct gctgaggcag
     4861 acagccgcca aagccgtgca acacttctac agtgtgaaac tgcagggcaa ggtgcccatg
     4921 cacaaactct tcctggagat gctggaggcc aaggtggtta acggcagcgg agagggcaga
     4981 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgagacgcgt
     5041 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     5101 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     5161 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     5221 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     5281 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     5341 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     5401 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     5461 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     5521 gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc
     5581 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     5641 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     5701 tactgcgacc tccctagcaa actggggcac aagctcaatt aaaagcttta aacgcgttga
     5761 gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgtaaaatt
     5821 catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt
     5881 gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc
     5941 attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact ttagcttgca
     6001 tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt actttctcta
     6061 atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac agttttagag
     6121 aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg
     6181 tggaaatatt cttattggta gaaacaacta caccctggtc atcatcctgc ctttctcttt
     6241 atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg
     6301 cccctctgct aaccatgttc atgccttctt ctctttccta cagctcctgg gcaacgtgct
     6361 ggttgttgtg ctgtctcatc attttggcaa aggattcact cctcaggtgc aggctgccta
     6421 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgttt
     6481 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     6541 ataaaggaaa tttattttca ttgcaatagt gtgttggaat ttctcgatcg ccgcggacac
     6601 gatccaagtg gagacctcca cagccaccag cctcgacttt tttcacaccc gcatcggggc
     6661 tctgagctgt cccagaagaa ggggtcttct tgcttcctgg ccatgtgcag actcctgggg
     6721 acagcagatg gggaggtggg gatggggagg gtaggggcgg ggggctcatc tgtcacccgc
     6781 attttctttg gaattttttt tttccttctc catgggcagt gctaaggctt gggccaggga
     6841 cgacttccct tagagctgga gaccaccaga ggaagcagcc ttcctgcaag ggatccattt
     6901 ctggacctct ccctatttag gacctggagg tatctggatg ggcagtgctt agtgcccggg
     6961 acccaagaga catagattgg gggctcctga aggtgttggt gtcacggtgg gcagtccctt
     7021 ggggcagaac gtctctgtgg cctatcccga ggctctgttc ctcctccatc tagctggctc
     7081 cctccacttt cccctttctt attgtcccag tacacccagt tctcagtgga tgctcctgct
     7141 agagtagcca catccccacc ccgaagaacc cctcccctgc ttcctgcccc tacctcagcc
     7201 agccggaact cactggctca gaaaagagtt gggttctgta cccactgctc tttttgcctg
     7261 ctgtttctcc ttctcctctt gggcatggcc agtctagaaa cctatggaga attcaggacc
     7321 tggccccacc agaggtcact tgagggactc tcaaggtcag tagcttaggt tggggtgtag
     7381 gatagcagag agacccagcg gtagagggaa agtctcattg cacctcggaa aggaaggagc
     7441 tctaaaggtc ccccttggcc cctcccttac ctcataaaaa ggcaaggctt ggctttgtgc
     7501 cataggaggg cagcctcctg gcatttgcac cagggatgtt tgatgcagcc ctcctgggtc
     7561 aattcccggc aagaccctgc agtctggttg tcgtataggg tgctccacca acccaccgca
     7621 gtcctaaaac ccctcagccc taggcacatg aatgcacctt tcacccaccc aaaaggcagt
     7681 cccaggctca tgctgtgtgt gtccttgggt ttgggttgac aggatgtcct aaagatgaag
     7741 cggttcattc tggaaaatgg acacgccact ttcatgacaa gtcctccggc tttacctgct
     7801 ctgatccctt cctggagtgc aagggtcccc tctttcaggc gttctggcac ctgccactac
     7861 tgctggctct gcctcccggg tgacacctct aggagactgg tcacttcaca gacttaatgc
     7921 taaacttccc cagagagcct ttccctgccc ttttgcccac aaaggggttc atgcctctgc
     7981 atgggctctc cttgcccatg atccgcggcc gccatggtca tagctgtttg acgtcaggtg
     8041 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     8101 atatgtatcc gctcacgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     8161 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     8221 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     8281 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     8341 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     8401 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     8461 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     8521 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     8581 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     8641 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     8701 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8761 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8821 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8881 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8941 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     9001 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     9061 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     9121 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     9181 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     9241 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     9301 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     9361 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     9421 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     9481 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     9541 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     9601 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     9661 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9721 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     9781 ggaaaaacgc cagcaacgcg gcctttttac ggttctggcc ttttgcacgg accgtagagt
     9841 cgagcagtgt ggttttcaag aggaagcaaa aagcctctcc acccaggcct ggaatgtttc
     9901 cacccaatgt cgagcagtgt ggttttgcaa gaggaagcaa aaagcct
//
