LOCUS       sequence-238913-        7055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for mouse Ddx42 to insert 2A-tdiRFP670-PEST into 
            immediately upstream of stop codon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7055)
  AUTHORS   Ochiai H, Hayashi T, Umeda M, Yoshimura M, Harada A, Shimizu Y, 
            Nakano K, Saitoh N, Liu Z, Yamamoto T, Okamura T, Ohkawa Y, Kimura 
            H, Nikaido I
  TITLE     Genome-wide kinetic properties of transcriptional bursting in mouse 
            embryonic stem cells.
  JOURNAL   Sci Adv. 2020 Jun 17;6(25):eaaz6699. doi: 10.1126/sciadv.aaz6699. 
            eCollection 2020 Jun.
  PUBMED    32596448
REFERENCE   2  (bases 1 to 7055)
  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: "Sci Adv."; 
            date: "2020-06-17"; volume: "6(25):eaaz6699. doi"; pages: " 
            10.1126/sciadv.aaz6699. eCollection 2020 Jun"
COMMENT     Sequence Label: pTV-Ddx42-tdiRFP670-PEST
FEATURES             Location/Qualifiers
     source          1..7055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1644..1697
                     /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"
     protein_bind    complement(1698..1731)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1740..1760
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3723..3842
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     protein_bind    3845..3878
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4868..4888)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4868..4886)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4907..4923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4907..4923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4920..4942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4931..4947
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4955..4961,4962..4979,4980..4985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5000..5021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5138..5155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5309..5897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5389..5408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6068..6859,6860..6928))
                     /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     6691..6710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6929..7019)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgc
      661 tatcccgaaa gttccctata ccctccctaa aataaacctt aaaaaaaaaa aaacaacaac
      721 aacaacttta gaacagtatc tcttggaaga gtgaggaaac cttcctgaga aagcagctga
      781 gtctgtaagc ttcatgctag atttagagcc ttatgttcac ttgattttca catacaggca
      841 ttaacagtat cctgcttgaa tacacaagtg gagtttctgt acactttatc attaacgatt
      901 atttgtaaca ttatttgtat attgaagttt gttatactta ttcctggtgt cttttttttt
      961 ttttctttca gtcccagtac aagagtcact ttgttgcagc cagtttaagt aatcagaagg
     1021 ccggaacctc ttctgcaggg gcaagtggat ggactagtgc aggaagctta aattctgttc
     1081 caactaattc agcacagcag ggccataata gtcctgacaa ccccatgacc agttctacca
     1141 agaacatccc aggcttcaac aactctggga atatcagtag tgccccagtg acctaccctt
     1201 ctattggagc ccaaggagtc aacaacacag cttcagggaa taacagccgt gaagggattg
     1261 ggggtggcaa tgggaagaga gagagatata ctgagaaccg gggtggtagc cgtcatagcc
     1321 atggagatgg tggcaatcgg catggagatg gtgggcgcca tggagatgga taccgttatc
     1381 cagaaagtgg tagccgtcat actgatggcc accgacatgg agagaccagg catggaggaa
     1441 gtgccggcag acacggagag agccggggtg caaatgatgg caggaatgga gaaagcagga
     1501 aagaaggttt taatcgtgaa aacaagatgg accccaaggt ggatagcagc aggatggaca
     1561 aagtggacag caagacagat aagacacctg atggctttgc tgttccagag ccacccaaaa
     1621 gaaagaaaag ccgatgggat agtgagggaa gaggtagcct gctaacttgc ggggacgtag
     1681 aggaaaatcc tggtcccata acttcgtata gcatacatta tacgaagtta tcaatgggcc
     1741 caaaaaagaa aagaaaagtt gaattcatgg ccagaaaagt ggacctgacc agttgtgacc
     1801 gagagccgat acacattcct ggctctatcc agccatgtgg gtgtttgctg gcctgcgatg
     1861 cccaggcagt caggattact cgaattaccg aaaatgcggg tgccttcttt ggccgtgaaa
     1921 cgccccgtgt cggcgaactc ttagccgatt atttcggaga aaccgaggct cacgcactta
     1981 ggaacgcact ggcgcaatca agcgatccca aacgccctgc tctcatcttt gggtggagag
     2041 atggactgac aggccgtact ttcgacataa gccttcaccg gcatgacgga accagtataa
     2101 ttgagtttga accggccgcc gctgagcaag cagacaaccc ccttcgcctg accagacaaa
     2161 ttatcgcccg cactaaggag ctgaaatcgc tggaagaaat ggccgctcgt gttcccaggt
     2221 atcttcaggc aatgctgggt tatcataggg taatgctcta cagattcgcc gacgatggct
     2281 ctggtatggt aataggggag gccaagcgat ccgacctgga gtcttttctg gggcagcatt
     2341 tccccgcatc tctagtaccc cagcaggcaa ggcttttgta cctcaagaat gccatccgcg
     2401 tggtatcgga ctcccgcggt attagtagca ggatcgttcc agaacacgac gcctctggag
     2461 ctgcgctgga tctgtctttc gctcacctcc gatctatcag cccttgtcat ttagagtttc
     2521 tgcgaaatat gggggtcagc gcaagtatgt ccctgagcat aataatcgat ggaacactat
     2581 ggggcctaat aatttgccat cattacgaac ctagagcagt accaatggca cagcgagtgg
     2641 ccgccgaaat gttcgccgat tttttgtctc ttcattttac tgccgctcac catcagcgcg
     2701 gccacgggac tggctcgaca ggttctggca gtagtgggac tgcttcatca gaaaataata
     2761 acatggccat ggccagaaaa gtggacctga ccagttgtga ccgagagccg atacacattc
     2821 ctggctctat ccagccatgt gggtgtttgc tggcctgcga tgcccaggca gtcaggatta
     2881 ctcgaattac cgaaaatgcg ggtgccttct ttggccgtga aacgccccgt gtcggcgaac
     2941 tcttagccga ttatttcgga gaaaccgagg ctcacgcact taggaacgca ctggcgcaat
     3001 caagcgatcc caaacgccct gctctcatct ttgggtggag agatggactg acaggccgta
     3061 ctttcgacat aagccttcac cggcatgacg gaaccagtat aattgagttt gaaccggccg
     3121 ccgctgagca agcagacaac ccccttcgcc tgaccagaca aattatcgcc cgcactaagg
     3181 agctgaaatc gctggaagaa atggccgctc gtgttcccag gtatcttcag gcaatgctgg
     3241 gttatcatag ggtaatgctc tacagattcg ccgacgatgg ctctggtatg gtaatagggg
     3301 aggccaagcg atccgacctg gagtcttttc tggggcagca tttccccgca tctctagtac
     3361 cccagcaggc aaggcttttg tacctcaaga atgccatccg cgtggtatcg gactcccgcg
     3421 gtattagtag caggatcgtt ccagaacacg acgcctctgg agctgcgctg gatctgtctt
     3481 tcgctcacct ccgatctatc agcccttgtc atttagagtt tctgcgaaat atgggggtca
     3541 gcgcaagtat gtccctgagc ataataatcg atggaacact atggggccta ataatttgcc
     3601 atcattacga acctagagca gtaccaatgg cacagcgagt ggccgccgaa atgttcgccg
     3661 attttttgtc tcttcatttt actgccgctc accatcagcg cggatccgcg tttaaactcg
     3721 agtctcacgg cttccctccc gaggtggagg agcaggccgc cggcaccctg cccatgagct
     3781 gcgcccagga gagcggcatg gatagacacc ctgctgcttg cgccagcgcc aggatcaacg
     3841 tcttgaagtt cctattctct agaaagtata ggaacttcta gtagagggta tgtgaaaatc
     3901 agcgtcgtca gtcgtcttta attttatttt tagaaagatt ttggtaatga ggcgtctcag
     3961 ggctgggttg gggtccaaga tatacagacc ccctgccctt ggtggagaac tggagagtgg
     4021 attgtacctt catctggtgt tcattcagag gtttccttgg gaacctgagc tggccctgga
     4081 aagcagagag ctgggaagct tttgtttgtt gtgggtgagt tttaaggagg gtaagtagaa
     4141 tacttaggga gaaagggcct tttagggcac aaaaactcac tctaggttta taatagctta
     4201 tattttttac taaaatgtca ccttataaac atctataaat tggacttttt ttcttagctg
     4261 tggccaggca gtcccaactt ctgccctgtt agggatgctt ggggagcagt ggtgctttca
     4321 gatgtttgca gggaagatgc ctgagcgctc aggttcccca gactgtttcc agccctgaga
     4381 cacgtaggaa agacagcctt ccagacacag gctttgaaat cctaagccat taggactgaa
     4441 ggaatgatca tcttggaagg attccaggat aatccaggcc tggagtactg ctaaatttac
     4501 catttaaaac caaccccaac agtagcattc aaagtgtcta tttgttttgt tttgtttttt
     4561 taaacttgcc ctaatagtag aaaaaagtct tctgctgaaa tgattttgat ttttgttcta
     4621 tcttgtttat aaaagaaaaa tatacaaact ttgaattttg tgatttttgt gaaggtttct
     4681 ttaagatgtg cattccctct ctgcttgctc gatagtaaat gtcttactac tgggacctga
     4741 gggctgtcgt ttctaccata caagttagtt gctacatatt aaattatggc ttggaacagt
     4801 acagcccatg atgtgaaatg tggggaagtg cttgcttatt aaactatcag agctccagct
     4861 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     4921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6181 ctggccccag tgctgcaatg ataccgcggc tgccacgctc accggctcca gatttatcag
     6241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6901 gggcgacacg gaaatgttga atactcatac tcttcctttt caatattatt gaagcattta
     6961 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7021 agggattccg cgcacatttc cccgaaaagt gccac
//
