LOCUS       sequence-328550-        8675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8675)
  AUTHORS   Nakamura T, Liu YJ, Nakashima H, Umehara H, Inoue K, Matoba S, 
            Tachibana M, Ogura A, Shinkai Y, Nakano T
  TITLE     PGC7 binds histone H3K9me2 to protect against conversion of 5mC to 
            5hmC in early embryos.
  JOURNAL   Nature. 2012 Jun 3;486(7403):415-9. doi: 10.1038/nature11093.
  PUBMED    22722204
REFERENCE   2  (bases 1 to 8675)
  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"; journalName: 
            "Nature"; date: "2012-06-3- 3"; volume: "486"; issue: "7403"; pages:
            "415-9"
COMMENT     Sequence Label: pcDNA4-FLAG-Jhdm2a-H1122A
FEATURES             Location/Qualifiers
     source          1..8675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             677..700
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4364..4393
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4409..4426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4449..4466)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4455..4679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4725..5153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4812..4831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5022..5043
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5162..5182)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5167..5496
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5347..5482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5409..5428
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5544..5591
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5610..5984
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6114..6235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6151..6170)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6205..6224
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6284..6300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6284..6300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6297..6319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6308..6324
                     /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(6332..6338,6339..6356,6357..6362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6377..6398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6515..6532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6686..7274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6766..6785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7445..8236,8237..8305))
                     /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     8068..8087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8306..8410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8485..8504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttatggact acaaagacga tgacgacaag ggatccatgt acaaatctct
      721 tctagacaaa gctggcttgg gagccataac ttctgttcgg tttcttggag atcaacaaag
      781 tgtatttgtt tccaaagacc ttttgaaacc tatacaggat gttaacagtc ttcggctttc
      841 ccttactgat aatcagacag tcagtaagga atttcaagct ttgattgtaa aacatttgga
      901 tgaaagccat cttttacaag gtgacaagaa ccttgttggt tcagaagtaa aaatttatag
      961 cttggaccca tctactcagt ggttttcagc aactgttgta catggaaacc catcatccaa
     1021 aactcttcaa gtcaactgtg aggagattcc agcactgaaa attgtcgacc cagcactgat
     1081 tcatgttgaa gttgtacatg acaactttgt gacatgtggt aattctacaa gaactggagc
     1141 tgtaaaacgc aagtcttctg agaataacgg aagttcggtt tctaaacaag caaaatcttg
     1201 ttctgaggcc tctcccagta tgtgtcctgt acagtctgtt cccacaacag tgtttaagga
     1261 gatcctgctt ggctgtactg cagcaactcc atctagcaag gacccaagac agcaaaatac
     1321 tccccaggca gccaattctc cacctaacat tggagcaaaa cttcctcaag gatgtcataa
     1381 gcagaactta ccagaagaac tttcttcctg tctaaacaca aaacctgaag taccgagaac
     1441 aaaaccagat gtctgcaaag aaggattact ttcttcaaaa tcttctcagg ttggagctgg
     1501 agacttgaaa attctgagtg agcccaaagg tagctgtatc cagcctaaaa caaacactga
     1561 tcaggagagc agactggagt ctgctccaca gccagtcact ggccttccaa aggagtgctt
     1621 gcctgcaaag acttcctcta aggcagaact ggacattgcc accactcctg aactgcagaa
     1681 gcatctagaa catgcagctt ccacatccga tgacctttca gataagccag aagtgaaagc
     1741 aggtgtcact agccttaata gttgtgcaga aaagaaggtc gaaccttcac atttaggttc
     1801 ccagtcacag aatttaaagg aaacttcagt aaaagtagat aatgaaagct gttgtacaag
     1861 aagcagtaat aaaacccaga ctcccccagc ccggaagtca gttttgacag acccagataa
     1921 agtcaggaag ctgcagcaga gcggagaggc ctttgttcag gatgactcct gtgttaacat
     1981 cgtggcacag ctgcccaagt gtcgggagtg tcgactagac agcctgcgca aggataagga
     2041 ccagcagaag gactctcctg tgttttgtcg ctttttccac ttcaggagat tacaattcaa
     2101 caagcatggt gtgttgcggg tagaaggctt cttaacacca aacaagtatg acagtgaagc
     2161 gattggcttg tggctgcctt tgaccaaaaa tgttgtgggg actgatttgg acacagcaaa
     2221 atatatcctg gccaatattg gagaccactt ctgtcaaatg gtgatttctg agaaggaagc
     2281 tatgtcgact attgagccac acagacaggt tgcttggaaa cgagctgtca aaggagttag
     2341 agaaatgtgt gatgtgtgtg acacaaccat tttcaacctg cactgggtgt gccctcggtg
     2401 tgggtttgga gtatgtgtag attgctaccg gatgaagagg aagaactgcc aacagggtgc
     2461 tgcctacaag actttctctt ggataaggtg tgtgaagagt cagatacatg agcctgagaa
     2521 cctgatgccc acacagatta ttcctggcaa agccctctac gatgttggag acattgtgca
     2581 ttctgtcaga gcaaaatggg gcataaaggc caattgtccc tgctccaaca ggcagttcaa
     2641 gctcttctca aagccagcct taaaggaaga cctgaaacag acatccttgt ctggagaaaa
     2701 accaactctt gggaccatgg tccagcaaag ttcccctgtt ttggagccag tggctgtgtg
     2761 cggggaagca gcctccaagc cagccagcag cgtgaagccc acctgtccca ccagcacttc
     2821 acctttaaac tggctagctg accttaccag tgggaatgtc aacaaggaga ataaggaaaa
     2881 acagctgact atgccaattt taaagaatga aatcaaatgc cttccaccct tgccccctct
     2941 gaacaagccc agcacagtcc tccatacttt taacagcacc atcttgacac ctgtgagcaa
     3001 caataattca ggtttcctta gaaatctttt gaattcatcc acagcaaaga cagaaaatgg
     3061 attgaaaaac acacccaaaa ttcttgatga catctttgcc tctttggtgc aaaacaagac
     3121 ttcttctgat tcatccaaga ggcctcaagg actgacaatc aagcctagca ttcttggctt
     3181 tgacactcct cactactggc tgtgtgacaa ccgcctgctg tgcttgcaag accccaacaa
     3241 taagagcaat tggaatgttt ttagggaatg ctggaaacaa gggcagccag tgatggtgtc
     3301 gggcgtgcat cataaattaa acactgaact ctggaaaccc gagtccttca gaaaagagtt
     3361 tggtgagcag gaagtagacc tagtcaattg taggaccaat gaaatcatca ctggagccac
     3421 agtcggagac ttctgggatg gatttgaaga tgttccaaac cgtttgaaaa acgacaaaga
     3481 aaaagaacca atggtgttga aacttaagga ctggccgcca ggagaagact tcagagacat
     3541 gatgccttcc aggtttgatg atctgatggc caacattcca ctgcctgagt acaccaggcg
     3601 agatggcaaa ctgaacctgg cttccagact gccaaactac tttgtacggc cagacctggg
     3661 ccccaagatg tacaatgctt atggattgat cactccagag gatcggaaat atgggaccac
     3721 aaatcttgcc ttagatgtat ctgatgcagc caatgtcatg gtttatgtgg gaattcccaa
     3781 aggacagtgt gaacaagaag aagaagtcct tagaaccatc caagatggag attctgatga
     3841 actcacaatc aagagattta ttgaaggaaa agagaagcca ggagcccttt ggcacatata
     3901 tgctgctaaa gacacagaga agataagaga attccttaaa aaggtatcag aggagcaggg
     3961 tcaagacaac cctgcagacc atgaccctat ccacgatcag agctggtatt tagaccgatc
     4021 gctgagaaag cgcctctatc aagagtacgg cgtgcaaggc tgggctattg tacagtttct
     4081 tggggatgtg gtgtttatcc cagcaggagc gccacatcag gttcataact tatacagctg
     4141 tatcaaagtg gctgaagact ttgtgtctcc agagcatgtt aaacactgct tctggcttac
     4201 tcaggaattc cgttacttgt cacagactca taccaaccat gaagataaat tgcaggtgaa
     4261 aaatgttatc taccatgcag tgaaagatgc agttgctatg ctgaaagcca gtgaatccag
     4321 tttgggcaaa ccttagcggc cgctcgagtc tagagggccc ttcgaacaaa aactcatctc
     4381 agaagaggat ctgaatatgc ataccggtca tcatcaccat caccattgag tttaaacccg
     4441 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4501 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4561 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4621 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4681 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4741 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4801 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4861 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4921 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4981 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5041 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5101 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     5161 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5221 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5341 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5401 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5461 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5521 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     5581 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     5641 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     5701 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     5761 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     5821 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     5881 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     5941 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     6001 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6061 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     6121 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6181 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6241 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     6301 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6361 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     7021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     7081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     7141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     7201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8401 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     8461 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8521 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8581 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     8641 gctgcttcgc gatgtacggg ccagatatac gcgtt
//
