LOCUS       sequence-100312-        7238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mRNA synthesis or expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7238)
  AUTHORS   Matoba S, Liu Y, Lu F, Iwabuchi KA, Shen L, Inoue A, Zhang Y
  TITLE     Embryonic Development following Somatic Cell Nuclear Transfer 
            Impeded by Persisting Histone Methylation.
  JOURNAL   Cell. 2014 Nov 6;159(4):884-95. doi: 10.1016/j.cell.2014.09.055. 
            Epub 2014 Oct 30.
  PUBMED    25417163
REFERENCE   2  (bases 1 to 7238)
  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.cell.2014.09.055"; journalName: "Cell"; date: "2014-11-6-
            6"; volume: "159"; issue: "4"; pages: "884-95"
COMMENT     Sequence Label: pcDNA-flag-mKdm4d(H189A)-polyA
FEATURES             Location/Qualifiers
     source          1..7238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             911..934
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2741..2770
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2786..2803
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2826..2843)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2832..3056
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3102..3530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3189..3208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3399..3420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3539..3559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3544..3874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3725..3860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3787..3806
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3941..4735
                     /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(3995..4014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4605..4624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4911..5032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4948..4967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5002..5021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5081..5097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5081..5097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5094..5116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5105..5121
                     /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(5129..5135,5136..5153,5154..5159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5174..5195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5312..5329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5483..6071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5563..5582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6242..7033,7034..7102))
                     /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     6865..6884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7103..7207)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttatg gactacaaag acgatgacga caagggatcc actagtaacg gccgccagtg
      961 tgctggaatt cacaagacga agtccacatg tgctcagaat ccaaattgca gcataatgat
     1021 atttcgtcca accaaagaag agtttaatga ttttgacaaa tacattgctt acatggagtc
     1081 ccaaggggca caccgagctg gactggccaa ggtcatccca ccaaaagaat ggagggccag
     1141 gcagtcttat gacaatatca gcaacatctt aatagcaact cccctgcagc aagtggtctc
     1201 tgggcaggca ggcgtgttca cccaatacca taagaagaag aaaggcatga cagtggggga
     1261 gtaccgtgag ctggccaaca gcaaaaagta ccagaccccg ccacacctgg attttgaaga
     1321 tttggagcga aaatactgga agaatcgcct gtatgagtca ccgatttatg gtgctgacgt
     1381 cagcggctcc ctgtttgatg ggaagactca acagtggaat gtgggtcacc tgggaacaat
     1441 tcaagaccta ttggaacagg aatgtggcat agtgattgag ggcgtcaaca cgccctacct
     1501 gtactttggc atgtggaaga ccacctttgc gtgggctacg gaggacatgg acctgtacag
     1561 tatcaactac ctacactttg ggcagcccaa gacctggtat gctgtgcccc ctgagcatgg
     1621 caggcgcctg gagcgcctgg ccagggaact cttccctggc agctcccagg gctgccaggc
     1681 cttcctgagg cacaaggtgg cgctcatctc gcccactgtg cttaaggaga atggcatccc
     1741 ctttggtcgc atcacccagg aggctgggga gttcatggtc acctttccct atggctacca
     1801 cgcgggcttc aaccatggct tcaactgcgc agaggccatc aattttgcca ccccaaggtg
     1861 gattgactat ggcaaggtgg catctcagtg cagctgtggg gaggccaggg tgagcttctc
     1921 catggacgcc tttgtgagga tcttgcagcc tgagcgatat gaactgtgga aacgtggcca
     1981 agatcaggca gttgtggacc acacagagac tatggtgtct accagtcagg agctcaccac
     2041 ccggcgggtg accaaagcac caagaaaaac ttggggcttg aagcgtctcc ggcttcgcca
     2101 ggtctcacga tctcttctgc ctatagccac ggtaagtaac gttccttgca acatgcaggt
     2161 gtgccacacc tccaggcaac catcagatgt gaaaggtgat gatgtccaga agtctgactc
     2221 agccagagcc tcaccacatc ctctgagtct gccttcttct ggtcacatgt caactagaag
     2281 atgtagtctt ggtcgtcgtc cttgtgaact aggagctcag gagtcctcca atggagctcc
     2341 agtcaagagg caacttccag caggcagaga tgacacaagc cccagtccag agcttcagcc
     2401 ccaggctgtg agtggagact taatagtcga ctcaggactt gtgaaccctg gcccacagca
     2461 tcttatgaca gcttctgaag ggggattgac ctccgacccc taagaattct gcagatatcc
     2521 atcacactgg cggccgctcg actatgggca ccaaagaacc tgtaaacgtt atctttttaa
     2581 attgaatgtg cacaaataaa agtttggaaa agaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2641 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaatacgc
     2701 atcatacgct gtggcgtacc tcgagtctag agggcccttc gaacaaaaac tcatctcaga
     2761 agaggatctg aatatgcata ccggtcatca tcaccatcac cattgagttt aaacccgctg
     2821 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2881 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2941 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3001 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3061 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3241 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     3301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     3601 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3721 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3781 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3841 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3901 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3961 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4021 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4081 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4141 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4201 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4261 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4321 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4381 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4441 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4501 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4561 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4621 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4681 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4741 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4801 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4861 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4921 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4981 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5041 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5101 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5161 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5221 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5281 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5341 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5401 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5461 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5521 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5581 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5641 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5701 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5761 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5821 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5881 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5941 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6001 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6061 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6121 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6181 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6241 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6301 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6361 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6421 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6481 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6541 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6601 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6661 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6721 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6781 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6841 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6901 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6961 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7021 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7081 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7141 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7201 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
