LOCUS       sequence-200685-        8934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mouse Kdm4b in mammalian cells. It has a T7 promoter for in 
            vitro transcription. Since polyA is encoded on the vector, 
            polyA-tailing is unnecessary..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8934)
  AUTHORS   Inoue A, Jiang L, Lu F, Zhang Y
  TITLE     Genomic imprinting of Xist by maternal H3K27me3.
  JOURNAL   Genes Dev. 2017 Oct 1;31(19):1927-1932. doi: 10.1101/gad.304113.117.
  PUBMED    29089420
REFERENCE   2  (bases 1 to 8934)
  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: 
            "10.1101/gad.304113"; journalName: "Genes Dev"; date: "2017-10-1- 
            1"; volume: "31"; issue: "19"; pages: "1927-1932"
COMMENT     Sequence Label: pcDNA-Flag-Kdm4b-pA
FEATURES             Location/Qualifiers
     source          1..8934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        483..686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     637..657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     731..750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        731..749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             779..802
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4306..4335
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4351..4368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4391..4408)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4397..4621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4667..5095
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4754..4773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4964..4985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5104..5124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5109..5438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5289..5424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5505..6299
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5559..5578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6169..6188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6475..6596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6512..6531)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6566..6585
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6645..6661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6645..6661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6658..6680)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6669..6685
                     /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(6693..6699,6700..6717,6718..6723))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6738..6759
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6876..6893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7047..7635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7127..7146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7806..8597,8598..8666))
                     /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     8429..8448
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8667..8771)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8846..8865)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
       61 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      721 ttatcgaaat taatacgact cactataggg agacccaagc tggctagtta agcttatgga
      781 ctacaaagac gatgacgaca agggatccac tagtaacggc cgccagtgtg ctggaattgg
      841 gtccgaggac cacagcgccc agaaccccag ctgtaagatc atgactttcc gccccaccat
      901 ggatgaattc agggacttca acagatatgt ggcgtacatt gagtcgcagg gtgcccaccg
      961 tgctggcctg gccaagatca ttccacccaa ggagtggaag ccacggcaga cgtatgatga
     1021 catcgacgat gtggtcatcc cagcgcctat tcagcaggtg gtgacgggcc agtcggggct
     1081 tttcacacag tacaacatcc agaagaaggc catgactgtg ggggagtacc gccgcctagc
     1141 caatagtgaa aagtactgta ccccacgcca tcaggacttc gatgacttgg agcgcaagta
     1201 ctggaagaac ctgacctttg tctcacccat ctatggagct gacatcagtg gctccctgta
     1261 cgacgatgac gtggcgcagt ggaacatcgg caatctgagg accatcctag acatggtgga
     1321 gcgcgaatgc gggaccatca ttgagggcgt gaatacaccc tacttgtact ttggcatgtg
     1381 gaagaccacg tttgcctggc acacagagga catggacctc tacagcatca actacctgca
     1441 cttcggcgag cctaagtcct ggtacgccat accaccggag cacggcaagc gcctggagcg
     1501 cctggccata ggcttcttcc ctgggagctc acagggctgt gatgccttcc taaggcacaa
     1561 gatgacactc atctcgccca tcatcctgaa gaagtacggc atccccttca gccggattac
     1621 acaggaagct ggggagttca tgatcacgtt cccctatggt taccatgcgg gcttcaacca
     1681 tggctttaac tgcgctgagt ccactaactt cgccacactg cgctggatcg actatggcaa
     1741 agtggccaca cagtgtacat gtcgaaagga catggtcaag atctccatgg acgtgtttgt
     1801 acgcatcctg cagcccgaac gctatgagca atggaagcaa ggccgtgacc tcacagtgct
     1861 ggaccacacg aggcccacag ccctgagcag tcctgagctc agctcctgga gtgcctcgcg
     1921 cacctccatc aaggccaagc tcctgcgcag acaaattagt gtgaaggaga gcagaccctg
     1981 gagaaaggct gaggaggaga ggaggcggga gccgaccagg aggccagggc cggcgtcaca
     2041 ccggagacgg agccagccca agaagtcaaa acctgaagag tccaggtctc caggggaagc
     2101 cacggctggg gtatctactc tggatgaggc caggggctgc tcccgaggag aggccatgcc
     2161 tgaggacgag gaggaggagg agctgctacc ttcacagggc catgaggccg agggtgtgga
     2221 agaggatggc agaggcaagc ctcggcctac taaggccaga aataagaaga agactcccag
     2281 cccgtcgtca cccccgttgc tgtctgcccc accagcccta ttccccaccg aggaggtcct
     2341 gaggccacca ccccaaccca agtctccagg gccagctatg ggccccatgg ctgcagaggg
     2401 gggtcctcca ccaacacccc tcaatgtcgt gcctcctggg gcgccagttg aggaagcaga
     2461 ggtgcggcca cggcccatta tcccaatgct gtacgtgttg ccccgcacca gcagcaccga
     2521 tggggacagg gagcactctg cccacgcaca gttggcaccc atggagctgg ggccagaaga
     2581 ggaaaaccag gcccaggctg gtgactcaca gggaacaacc cctttctcta agctgaaggt
     2641 ggagatcaaa aagagccggc gtcacccctt gggtcggcca cccactcgat ccccgctgtc
     2701 tgtggtcaag caggaggcct caagtgatga agaagccttc ctgttctcag gagaggatga
     2761 tgtcactgat cctgaggcct tgagatcgct gctgtccctg cagtggaaga acaaggcggc
     2821 cagcttccaa gccgagagga agttcaatgc ggcagccgcc ctcagtgagc cctactgtgc
     2881 catctgtacc ctcttctacc cctacagcca gtcagtacag acagagcggg acagtgctgt
     2941 ccagccgccc tccaaaagtg gccagaggac acggccactg atccctgaga tgtgcttcac
     3001 ttcaagtggg gagaacacag agccgctgcc tgctaactcc tacgtcggtg aggacggcac
     3061 cagtccactc atctcctgtg cccactgctg cctgcaggtc catgccagtt gctatggtgt
     3121 ccggcctgag ttggccaaag agggctggac atgttcccgg tgtgcggccc acgcttggac
     3181 tgcggaatgc tgtttgtgca acctgcgggg gggagcgcta cagaggacca cagagcacag
     3241 gtggattcac gtgatctgcg ccattgcagt ccctgaggta cgattcctca atgtgattga
     3301 acgcaaccca gtggacgtca gtgccatccc tgagcagcga tggaaactga aatgcattta
     3361 ctgccgcaag cgtatgaaga gagtgtcggg tgcctgcatc cagtgctcct atgagcactg
     3421 ctccacatcc ttccatgtga cctgtgcgca tgctgctggt gtcctcatgg aacccgatga
     3481 ctggccctat gtggtgtcca tcacctgcct caagcaccgg gccagcggag cagggggcca
     3541 gctcctgcgc acagtgtccc tgggccagat cgtcatcacc aagaaccgca atgggctcta
     3601 ctatcgctgc cgggtcattg gcaccactgc gcagacattc tacgaggtca actttgacga
     3661 tggctcctac agtgacaacc tgtacccaga gagcatcacg agcagagact gcctacggct
     3721 agggccacct cctgagggcg agctggtgga gcttcggtgg acagacggta atctgtacag
     3781 agccaggttt atctccatgg ccaccagcct catctaccag gtagagtttg aggatgggtc
     3841 tcagctgact gtgaagcgtg gggacatctt cacactggaa gaggagctgc ccaagagggt
     3901 gcgttcccgg ctgtcactga gcactggcac accacaggag cccagcttct ctggagacga
     3961 tgtgaaggct gccaagcgcc cacgggtggc ctcagtgctg gccaccacca ctgaggacac
     4021 agggcgtagc ccagaatacc tgtccttcat ggagagcctg ctgcaggcac agggccggcc
     4081 tggagcaccc ttctagggcc gctcgactat gggcaccaaa gaacctgtaa acgttatctt
     4141 tttaaattga atgtgcacaa ataaaagttt ggaaaagaaa aaaaaaaaaa aaaaaaaaaa
     4201 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4261 tacgcatcat acgctgtggc gtacctcgag tctagagggc ccttcgaaca aaaactcatc
     4321 tcagaagagg atctgaatat gcataccggt catcatcacc atcaccattg agtttaaacc
     4381 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4441 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4501 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4561 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4621 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4681 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4741 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4801 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4861 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4921 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4981 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5041 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     5101 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5161 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5281 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5341 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5401 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5461 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5521 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5581 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5641 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5701 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5761 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5821 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5881 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5941 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6001 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     6061 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6121 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6181 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6241 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6301 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6361 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6421 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6481 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6541 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6601 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6661 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6721 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6781 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7801 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7861 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7921 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7981 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8041 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8101 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8161 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8221 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8281 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8341 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8401 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8461 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8521 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8581 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8641 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8701 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8761 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8821 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8881 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa attt
//
