LOCUS       sequence-238329-        7127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for mouse Hdac1 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 7127)
  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 7127)
  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-mHdac1-tdiRFP670-PEST2
FEATURES             Location/Qualifiers
     source          1..7127
                     /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             1600..1653
                     /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(1654..1687)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1696..1716
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3679..3798
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     protein_bind    3801..3834
                     /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(4940..4960)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4940..4958)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4979..4995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4979..4995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4992..5014)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5003..5019
                     /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(5027..5033,5034..5051,5052..5057))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5072..5093
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5210..5227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5381..5969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5461..5480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6140..6931,6932..7000))
                     /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     6763..6782
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7001..7091)
                     /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 tgggtacccc
      661 ccttgctctc ataccaatgt ttctctctct gctgcctccc agtctgctcc tctgacaaac
      721 gcattgcctg tgaggaagag ttctcggact cagatgagga gggagaaggt ggtcgcaaga
      781 actcttctaa cttcaaaaaa gccaaaagag ttaaaacaga ggatgagaaa gagaaagatc
      841 ctgaagagaa aaaaggtgag tttggattag atcaggaagg agctgaccca gacccagcca
      901 gctcctttgg agcttaaact ggaggtttga gtcttgagtc tttcagtggc tcctagaggt
      961 ggtgagaacc acagaacagt gacagtgaca ctcttggtct ctgctggctg actgaccctg
     1021 gctcacatac cacccaagcc tttttcccag tttcccaggg ggctgcctct gcctgtgttc
     1081 ttgctgaaac aggctgggaa accggtctga cctgttttca gccaacctca ccagaggacc
     1141 tgggataggc ttttctcatt tttctataac cagtgaactg tggtattgaa gaaggggttt
     1201 caggatagag gagtccgact acctgagaag ccttcaagca tacctgttct cttattttct
     1261 agaagtcaca gaagaagaga aaaccaagga ggagaagcca gaagccaaag ggtgaggaaa
     1321 ggggctggtg gctctctgta gcactagagc atcatcacct tctgtgtcct acctctgtga
     1381 gagggaagga agagcaggga cagctgctgg tccagcctac agagagaggc ctctttaagg
     1441 gtggagtctg tatgtctctg cccagttccc agcagtcacc tgggcagcca gggtctatgg
     1501 aagtcactat agccatgttt gtgcttcagg gccactagca tgagttttct catgcctgtc
     1561 tgttctctct gcagggtcaa tgaggaagtt aaactggccg agggaagagg tagcctgcta
     1621 acttgcgggg acgtagagga aaatcctggt cccataactt cgtatagcat acattatacg
     1681 aagttatcaa tgggcccaaa aaagaaaaga aaagttgaat tcatggccag aaaagtggac
     1741 ctgaccagtt gtgaccgaga gccgatacac attcctggct ctatccagcc atgtgggtgt
     1801 ttgctggcct gcgatgccca ggcagtcagg attactcgaa ttaccgaaaa tgcgggtgcc
     1861 ttctttggcc gtgaaacgcc ccgtgtcggc gaactcttag ccgattattt cggagaaacc
     1921 gaggctcacg cacttaggaa cgcactggcg caatcaagcg atcccaaacg ccctgctctc
     1981 atctttgggt ggagagatgg actgacaggc cgtactttcg acataagcct tcaccggcat
     2041 gacggaacca gtataattga gtttgaaccg gccgccgctg agcaagcaga caaccccctt
     2101 cgcctgacca gacaaattat cgcccgcact aaggagctga aatcgctgga agaaatggcc
     2161 gctcgtgttc ccaggtatct tcaggcaatg ctgggttatc atagggtaat gctctacaga
     2221 ttcgccgacg atggctctgg tatggtaata ggggaggcca agcgatccga cctggagtct
     2281 tttctggggc agcatttccc cgcatctcta gtaccccagc aggcaaggct tttgtacctc
     2341 aagaatgcca tccgcgtggt atcggactcc cgcggtatta gtagcaggat cgttccagaa
     2401 cacgacgcct ctggagctgc gctggatctg tctttcgctc acctccgatc tatcagccct
     2461 tgtcatttag agtttctgcg aaatatgggg gtcagcgcaa gtatgtccct gagcataata
     2521 atcgatggaa cactatgggg cctaataatt tgccatcatt acgaacctag agcagtacca
     2581 atggcacagc gagtggccgc cgaaatgttc gccgattttt tgtctcttca ttttactgcc
     2641 gctcaccatc agcgcggcca cgggactggc tcgacaggtt ctggcagtag tgggactgct
     2701 tcatcagaaa ataataacat ggccatggcc agaaaagtgg acctgaccag ttgtgaccga
     2761 gagccgatac acattcctgg ctctatccag ccatgtgggt gtttgctggc ctgcgatgcc
     2821 caggcagtca ggattactcg aattaccgaa aatgcgggtg ccttctttgg ccgtgaaacg
     2881 ccccgtgtcg gcgaactctt agccgattat ttcggagaaa ccgaggctca cgcacttagg
     2941 aacgcactgg cgcaatcaag cgatcccaaa cgccctgctc tcatctttgg gtggagagat
     3001 ggactgacag gccgtacttt cgacataagc cttcaccggc atgacggaac cagtataatt
     3061 gagtttgaac cggccgccgc tgagcaagca gacaaccccc ttcgcctgac cagacaaatt
     3121 atcgcccgca ctaaggagct gaaatcgctg gaagaaatgg ccgctcgtgt tcccaggtat
     3181 cttcaggcaa tgctgggtta tcatagggta atgctctaca gattcgccga cgatggctct
     3241 ggtatggtaa taggggaggc caagcgatcc gacctggagt cttttctggg gcagcatttc
     3301 cccgcatctc tagtacccca gcaggcaagg cttttgtacc tcaagaatgc catccgcgtg
     3361 gtatcggact cccgcggtat tagtagcagg atcgttccag aacacgacgc ctctggagct
     3421 gcgctggatc tgtctttcgc tcacctccga tctatcagcc cttgtcattt agagtttctg
     3481 cgaaatatgg gggtcagcgc aagtatgtcc ctgagcataa taatcgatgg aacactatgg
     3541 ggcctaataa tttgccatca ttacgaacct agagcagtac caatggcaca gcgagtggcc
     3601 gccgaaatgt tcgccgattt tttgtctctt cattttactg ccgctcacca tcagcgcgga
     3661 tccgcgttta aactcgagtc tcacggcttc cctcccgagg tggaggagca ggccgccggc
     3721 accctgccca tgagctgcgc ccaggagagc ggcatggata gacaccctgc tgcttgcgcc
     3781 agcgccagga tcaacgtctt gaagttccta ttctctagaa agtataggaa cttctagtga
     3841 gcaaggtctg cagccccatc ttctccccaa gttcctcact tctcagattt tatattttct
     3901 attcctctgt gtatttatat aaaatatatt aactataacg tccccaggga ccagagcaca
     3961 gggcagtggt gctaggaggg ctcttcccag agcgaccttg tccatcctgc cacccccctc
     4021 cccccatttg cgtggaaaga aaacaaccag aagagaaaaa tcctgaactg ccaagtgcct
     4081 gcttaggagc tctgctaaga tgcccctgtt aaacttttta ggaggggtct gggtctccta
     4141 ggaccccttt tcaggcttgg gtaatagcag ccatttttag attggttctg tttatacctt
     4201 cccacccatc ttgggcaaac aaaaaaactt atattgcctg tcttccccca attctgtagg
     4261 tggaggttga tagcctagct tcctttttga gatattttca ttttgtgaaa ctctttgtaa
     4321 taaaacagct catttctata ccctcggccc tggttggttt aagttcttcc ccgtctcttg
     4381 tggggcgtca cttgcagacc tacctgcctt tgtcctccag gtaggtgcag tcctcttacc
     4441 cacaccccag aagaccactc aatgaaacat cactaaaggg acgacattta ttccttttcc
     4501 aaatgttaca gtaaatccgg gtggaagaat ggttttagca gttagaaaaa aaaaagtaca
     4561 aatctggggt ttggccatta aaagttattt acaacaatgg gagggggaaa aaaaaagaca
     4621 acaagaagtt gtttcacatt acagacctcc ctccctcctg aagcctaaca cttgcttatc
     4681 aagtcagaga catggttgat ttcacaagct ggaggtttga acttgagtaa gacatttata
     4741 aaaacccaga tggggcaatg tcctccccag cccaggtgcc actaggcaca gcacaagaga
     4801 ctaagaattc aacaggggaa aggctggaca ctcagggttt gggagtataa gcccccactt
     4861 ctggctcagg gggttgagaa gcaggataaa caaaacgaac ttggcggccg ccaccgcggt
     4921 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4981 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5041 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5101 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5161 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5221 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5281 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5341 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5401 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5461 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5521 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5581 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5641 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5701 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5761 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5821 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5881 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5941 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6001 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6061 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6121 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6181 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6241 agggcttacc atctggcccc agtgctgcaa tgataccgcg gctgccacgc tcaccggctc
     6301 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6361 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6421 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6481 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6541 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6601 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6661 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6721 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6781 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6841 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6901 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6961 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt ttcaatatta
     7021 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7081 aaataaacaa atagggattc cgcgcacatt tccccgaaaa gtgccac
//
