LOCUS       sequence-338401-        8322 bp    DNA     circular SYN 14-APR-2025
DEFINITION  full-length wt Avi-Mta2-3XFLAG-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 8322)
  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 8322)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-AviMta2-3xFiBsd
FEATURES             Location/Qualifiers
     source          1..8322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          656..1671
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1595..1617
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1679..1698
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1741..1785
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1783..1803
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3811..3876
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3919..4493,4494..4496,4497..4505)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4086..4103)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4313..4332
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4506..4904
                     /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(4936..4953)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4942..5166
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5213..5229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5213..5229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5226..5248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5237..5253
                     /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(5261..5267,5268..5285,5286..5291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5306..5327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5386..5581
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5432..5567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5587..5721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5789..5806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5960..6548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6040..6059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6719..7510,7511..7579))
                     /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     7342..7361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    8172..8306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        join(8320..8322,1..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 ggggggggns gcgcgccrgg sggggsgggg sggggsgrgg ggsggggsgg
      541 ggsgrggcgg agaggkgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
      601 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
      661 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
      841 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
      901 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     1021 tgccccgcgg tgcggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     1081 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     1321 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     1381 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     1441 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1621 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga catggctggt
     1741 ggcctgaatg acatctttga ggcccagaag atcgagtggc atgagaacct gtacttccag
     1801 ggagctgcta acatgtaccg ggtgggagat tacgtctatt ttgagaactc ctccagcaat
     1861 ccttacctgg ttagacggat tgaggagctc aataagactg caaatggaaa tgtggaagca
     1921 aaggttgtct gtcttttccg gcgaagggat atttctagta gcctcaacag cttggctgac
     1981 agtaatgctc gggagtttga ggaggaatca aaacagccag gggtctccga gcagcagaga
     2041 catcaactga agcaccgaga gctttttctt tctcggcagt ttgaatcatt accagccacc
     2101 cacataaggg ggaagtgcag cgtgactctc ttgaacgaga cagacatctt aaaccagtat
     2161 ctagacaagg aggactgctt tttttactca ctggtgtttg atcctgtgca gaagacactt
     2221 ctggctgatc aaggggagat cagagttggt tgcaaattcc aagctgagat cccagatcgt
     2281 ttggcagagg gagaatcaga taatcgaaac caacagaaga tggagatgaa agtctgggac
     2341 ccagacaacc ctctcacaga ccggcagatt gatcagtttc tcgtggtggc ccgagctgtg
     2401 ggaaccttcg cgagagccct agactgcagc agctcaattc ggcagccaag cttgcacatg
     2461 agtgcagctg ctgcgtcccg agatatcact ctgttccatg ccatggatac cttgcaaagg
     2521 aatggctatg acttggccaa ggctatgtca accctagtac cacagggagg cccggtgctc
     2581 tgtcgggatg agatggagga atggtctgct tctgaagcca tgctgtttga agaggctctg
     2641 gagaagtacg ggaaggattt caatgatatc cgccaggact ttctaccctg gaagtcactt
     2701 gcaagcatag tccagtttta ttacatgtgg aaaaccacag accggtatat tcagcagaaa
     2761 agattgaaag ctgctgaagc agacagcaaa ctgaaacagg tctacatccc cacttatacc
     2821 aagccaaacc ctaaccagat catttctgtg ggttcaaaac ctggcatgaa tggggctgga
     2881 ttccagaagg gcctgacttg tgagagctgc cacaccacac agtctgccca gtggtatgcc
     2941 tggggacccc ctaacatgca gtgccgactt tgtgcttcct gttggattta ttggaagaaa
     3001 tacgggggac tgaagacccc tacccaactt gaaggggctg ctcggggcac cacagagcca
     3061 cactcaagag gtcatttatc cagacctgaa gcccaaagtc tctcccccta tacaaccagt
     3121 gctaaccggg ccaagttgct ggctaagaac aggcaaactt tcttgctcca gaccacaaag
     3181 ctaactcgtc tagccagacg aatgtgccga gatctattac agccaaggag ggctgcccga
     3241 agaccctatg cacctatcaa tgccaatgcc atcaaggcag agtgctcgat ccgacttcct
     3301 aaggctgcca agacgccact gaaaattcac cctctggtgc ggctgccctt ggcaactatc
     3361 gtcaaagatc tggtggctca ggcacctctg aaaccaaaaa cacctcgggg taccaagaca
     3421 ccaatcaaca gaaaccagct gacccagaat cgaggcctgg ggggcattat ggtgaaacgt
     3481 tcctatgaga ctatggcagg ggcaggggtc cccttctctg ccaatggaag gcctctggct
     3541 tcagggattc gttcaagctc acagccagca gccaagcgtc agaaactaaa ccctgctgat
     3601 gctcccaatc cagtggtatt tgtggccaca aaggacacca gggctttaag gaaagctcta
     3661 actcacctgg aaatgcgccg agcggcccga aggcccaacc tacccctgaa ggtgaaacca
     3721 acgctgatga cagtgcggcc cccagttcct ctgcccgctt cttcacatcc tgccagcacc
     3781 aatgagccca ttgtcctgga ggatagcggc gactacaaag accatgacgg tgattataaa
     3841 gatcatgaca tcgactacaa ggatgacgat gacaagtagc ggccgctcga taagcttgat
     3901 atcgaattcc gccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg
     3961 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     4021 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     4081 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     4141 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     4201 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     4261 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     4321 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     4381 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     4441 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     4501 caaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     4561 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     4621 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgcgcagaac
     4681 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     4741 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gttcttctcg
     4801 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     4861 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaaatgcat cgatgatcta
     4921 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4981 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5041 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5101 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     5161 ctatggcttc tgaggcggaa agaacctgca gcccaagctt ggcgtaatca tggtcatagc
     5221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     5401 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5461 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5521 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5581 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5641 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5701 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5761 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7561 ggaaatgttg aatactcata ctcttccttt ttcaatgacc gctttggaaa aacaaagact
     7621 gtatttcctg gaaattaatg tttattcaat aaactgtgta ttcagctata ttcacatagt
     7681 ggtgaggctg aaatgaggcg ggaagaggcg gttggggctt aattatatca atttgggtgg
     7741 ccccacagcg cctccaaggc gccagtcctg ttttgacaag ttgcctctgg aagcctctct
     7801 tctttttctc cagagtaagc ggaggccagg ggcccccggc ctctgcttaa tactaaaaaa
     7861 aacagctgtt gtcatagtaa tgattgggtg gaaacattcc aggcctgggt ggagaggctt
     7921 tttgcttcct cttgcaaaac cacactgaca ttccaggcct gggtggagag gctttttgct
     7981 tcctcttgca aaaccacact gccctctgga gggcagttgc ctagcaacta attaaaagag
     8041 gatgtcgcac ggccagctgc ggtcagttag tcacttcctg cttaactgac ttgacatttt
     8101 ctattttaag agtcgggagg aaaattactg tgttggaggc cctccgccat cttctgaagc
     8161 tgaatcgaat taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     8221 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     8281 tcaatgtatc ttatcatgtc tggatctgat atcatcgtcg ac
//
