LOCUS       sequence-338420-        7858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full-length wt Avi-Mta3-3XFLAG-iBsd cDNA w/CAG promoter and 
            Blasticidin resistance, cloned from mouse ES cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7858)
  AUTHORS   Burgold T, Barber M, Kloet S, Cramard J, Gharbi S, Floyd R, 
            Kinoshita M, Ralser M, Vermeulen M, Reynolds N, Dietmann S, Hendrich
            B
  TITLE     The Nucleosome Remodelling and Deacetylation complex suppresses 
            transcriptional noise during lineage commitment.
  JOURNAL   EMBO J. 2019 Jun 17;38(12). pii: embj.2018100788. doi: 
            10.15252/embj.2018100788. Epub 2019 Apr 29.
  PUBMED    31036553
REFERENCE   2  (bases 1 to 7858)
  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: "EMBO J."; 
            date: "2019-06-17"; volume: "38(12). pii: embj.2018100788. doi"; 
            pages: " 10.15252/embj.2018100788. Epub 2019 Apr 29"
COMMENT     Sequence Label: pCAG-AviMta3-3xFiBsd
FEATURES             Location/Qualifiers
     source          1..7858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        379..656
                     /label=chicken beta-actin promoter
     intron          657..1672
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1596..1618
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1680..1699
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1742..1786
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1784..1804
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3347..3412
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3455..4029,4030..4032,4033..4041)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3622..3639)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3849..3868
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4042..4440
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(4472..4489)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4478..4702
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4749..4765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4749..4765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4762..4784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4773..4789
                     /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(4797..4803,4804..4821,4822..4827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4842..4863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4922..5117
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4968..5103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5123..5257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5325..5342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5496..6084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5576..5595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6255..7046,7047..7115))
                     /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     6878..6897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7708..7842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        join(7856..7858,1..377)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
      421 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
      481 gggcgggggg gggggggggn cgcgcgccrg gsggggsggg gsggggsgrg gggsggggsg
      541 gggcgaggcg gagaggkgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      601 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      661 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      721 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      781 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      841 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
      961 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
     1021 gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     1081 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     1141 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     1201 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     1261 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     1321 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     1381 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     1561 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     1621 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     1681 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcctcg acatggctgg
     1741 tggcctgaat gacatctttg aggcccagaa gatcgagtgg catgagaacc tgtacttcca
     1801 gggagcggcc aacatgtacc gggtcggaga ctacgtttat tttgagaatt cctccagcaa
     1861 cccgtacctc atcagacgga tagaggagct caacaagact gcaagtggca atgtggaagc
     1921 aaaagtcgtc tgcttttaca gaagacggga catctccaac acgcttatca tgctcgccga
     1981 caagcatgct aaagaaactg aggaggagtc ggagacgcca gttgaggccg acttgacgga
     2041 gaagcagaag caccagctga aacacaggga gctctttctg tcgcgccagt acgagtcact
     2101 gcctgcgaca catatcaggg ggaagtgcag cgtggccctg ctgaatgaga cagaatccgt
     2161 gttgtcatac cttgacaagg aggatacctt cttctactca ctggtatacg acccttcagt
     2221 gaaaacatta ttggctgaca aaggtgaaat cagagtgggc ccaaagtacc aagctgacat
     2281 tccagacatg ctaccagaag gagactcaga tgagagggaa caatcaaaat tggaagttaa
     2341 ggtttgggac cccaatagcc cacttacgga tcgacagatt gaccagtttt tagttgtagc
     2401 ccgtgccgtg ggaacatttg cccgagccct ggactgcagc agctccgtga ggcaacccag
     2461 ccttcacatg agtgcagccg cagcctcccg agacatcacc ttgtttcatg ccatggacac
     2521 actgtacagg cacggctatg acctcagcag cgccatcagt gtgctggtgc cgctcggagg
     2581 gccagtcctc tgcagggacg agatggagga gtggtcggcc tctgaagcct gcctattcga
     2641 agaagcactg gaaaaatacg gcaaagattt caacgacatc cgtcaggact ttctcccgtg
     2701 gaaatccttg actagcatca ttgaatatta ttacatgtgg aaaactactg acagatacgt
     2761 gcagcagaag cgcctaaagg ccgcagaagc cgagagcaaa ctgaaacaag tgtacatccc
     2821 gacttacaaa ccaaatccca accaaatctc cagcagcaac ggcaaggctg gcactgtgaa
     2881 tggagctgtg gggacccagt ttcagcccca gagcgccctc ctagggcgag cctgcgagag
     2941 ctgctacgcc acacagtctc atcagtggta ttcctggggc ccacctaaca tgcagtgtag
     3001 actctgcgcg acctgttggc tgtattggaa aaagtatgga ggtctgaaaa tgcctaccca
     3061 gtcagacgaa gagaagtctc ccagcccgac tgcagaggac ccgcgcgcga gaagccacat
     3121 gtcccggcag gccttgcagg gcatgccggt ccggaacacc ggaagcccca agtctgcagt
     3181 gaagacccgc caagctttct tccttcgtac tacgtatttc acaaaaattg ctcgtcaggt
     3241 ctgcaaaagc accctgcggc tgcggcaggc agcgagacgg ccgtttgttg ctattaatta
     3301 tgctgccatt agggcagaat gtaagacgct tttcaattcg agcggcgact acaaagacca
     3361 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc
     3421 gctcgataag cttgatatcg aattccgccc cccccccctc tccctccccc ccccctaacg
     3481 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3541 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3601 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3661 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     3721 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3781 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3841 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3901 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3961 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     4021 gatgataata tggccacaac catggccaag cctttgtctc aagaagaatc caccctcatt
     4081 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     4141 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     4201 ggaccttgcg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     4261 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     4321 cgacaggttc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     4381 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     4441 atgcatcgat gatctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     4501 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4561 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4621 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4681 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa cctgcagccc aagcttggcg
     4741 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4801 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     4921 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4981 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5041 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5101 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     5161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5221 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     5281 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5341 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5401 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5461 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5521 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5581 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5641 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5701 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5761 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5821 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5881 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5941 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6001 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6061 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6121 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6181 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6241 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6301 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6361 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6421 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6481 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6541 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6601 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6661 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6721 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6781 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6841 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6901 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6961 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7021 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7081 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atgaccgctt
     7141 tggaaaaaca aagactgtat ttcctggaaa ttaatgttta ttcaataaac tgtgtattca
     7201 gctatattca catagtggtg aggctgaaat gaggcgggaa gaggcggttg gggcttaatt
     7261 atatcaattt gggtggcccc acagcgcctc caaggcgcca gtcctgtttt gacaagttgc
     7321 ctctggaagc ctctcttctt tttctccaga gtaagcggag gccaggggcc cccggcctct
     7381 gcttaatact aaaaaaaaca gctgttgtca tagtaatgat tgggtggaaa cattccaggc
     7441 ctgggtggag aggctttttg cttcctcttg caaaaccaca ctgacattcc aggcctgggt
     7501 ggagaggctt tttgcttcct cttgcaaaac cacactgccc tctggagggc agttgcctag
     7561 caactaatta aaagaggatg tcgcacggcc agctgcggtc agttagtcac ttcctgctta
     7621 actgacttga cattttctat tttaagagtc gggaggaaaa ttactgtgtt ggaggccctc
     7681 cgccatcttc tgaagctgaa tcgaattaac ttgtttattg cagcttataa tggttacaaa
     7741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     7801 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatatca tcgtcgac
//
