LOCUS       sequence-497795-        7707 bp    DNA     circular SYN 20-JAN-2026
DEFINITION  Expresses mouse MyoD-K/R-ER(T)-3xT7 cDNA where K99, K102, and K104 
            residues of MyoD proteins were mutated to R..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  TITLE     ""Acute interventions reveal dynamic interplay between transcription
            factors and chromatin modifiers on enhancers""
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 20, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pWZLhygro-MyoD-K/R-ER(T)-3xT7
FEATURES             Location/Qualifiers
     source          1..7707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2124..2585
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2180..2197)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2407..2426
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2590..3615
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2590..3615
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             3804..4273
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4403..4420)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4574..5162)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4654..4673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5322..6113,6114..6182))
                     /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     5945..5964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6183..6287)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6318..6787
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6852..7051
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7252..7668
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     7598..7620
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     7636..7652
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 atggagcttc tatcgccgcc actccgggac atagacttga caggccccga cggctctctc
       61 tgctcctttg agacagcaga cgacttctat gatgatccgt gtttcgactc accagacctg
      121 cgcttttttg aggacctgga cccgcgcctg gtgcacgtgg gagccctcct gaaaccggag
      181 gagcacgcac acttctctac tgcggtgcac ccaggcccag gcgctcgtga ggatgagcat
      241 gtgcgcgcgc ccagcgggca ccaccaggcg ggtcgctgct tgctgtgggc ctgcagggcg
      301 tgcaggcgca ggaccaccaa cgctgatcgc cgcaaggccg ccaccatgcg cgagcgccgc
      361 cgcctgagca aagtgaatga ggccttcgag acgctcaagc gctgcacgtc cagcaacccg
      421 aaccagcggc tacccaaggt ggagatcctg cgcaacgcca tccgctacat cgaaggtctg
      481 caggctctgc tgcgcgacca ggacgccgcg ccccctggcg atcgaaatga aatgggtgct
      541 tcaggagaca tgagggctgc caacctttgg ccaagccctc ttgtgattaa gcacactaag
      601 aagaatagcc ctgccttgtc cttgacagct gaccagatgg tcagtgcctt gttggatgct
      661 gaaccgccca tgatctattc tgaatatgat ccttctagac ccttcagtga agcctcaatg
      721 atgggcttat tgaccaacct agcagatagg gagctggttc atatgatcaa ctgggcaaag
      781 agagtgccag gctttgggga cttgaatctc catgatcagg tccaccttct cgagtgtgcc
      841 tggctggaga ttctgatgat tggtctcgtc tggcgctcca tggaacaccc ggggaagctc
      901 ctgtttgctc ctaacttgct cctggacagg aatcaaggta aatgtgtgga aggcatggtg
      961 gagatctttg acatgttgct tgctacgtca agtcggttcc gcatgatgaa cctgcagggt
     1021 gaagagtttg tgtgcctcaa atccatcatt ttgcttaatt ccggagtgta cacgtttctg
     1081 tccagcacct tgaagtctct ggaagagaag gaccacatcc accgtgtcct ggacaagatc
     1141 acagacactt tgatccacct gatggccaaa gctggcctga ctctgcagca gcagcatcgc
     1201 cgcctagctc agctccttct cattctttcc catatccggc atatgagtaa caaacgcatg
     1261 gagcatctct acaacatgaa atgcaagaac gtggtacccc tctatgacct gctcctggag
     1321 atgttggatg cccaccgcct tcatgcccca gccagtcgca tgggagtgcc cccagaggag
     1381 cccagccaga cccagctggc caccaccagc tccacttcag cacattcctt acaaacctac
     1441 tacatacccc cggaagcaga gggcttcccc aacacgatcg ccgctgcctt ctacgcacct
     1501 ggaccgctgc ccccaggccg tggcagcgag cactacagtg gcgactcaga cgcgtccagc
     1561 ccgcgctcca actgctctga tggcatgatg gattacagcg gccccccaag cggcccccgg
     1621 cggcagaatg gctacgacac cgcctactac agtgaggcgg tgcgcgagtc caggccaggg
     1681 aagagtgcgg ctgtgtcgag cctcgactgc ctgtccagca tagtggagcg catctccaca
     1741 gacagccccg ctgcgcctgc gctgcttttg gcagatgcac caccagagtc gcctccgggt
     1801 ccgccagagg gggcatccct aagcgacaca gaacagggaa cccagacccc gtctcccgac
     1861 gccgcccctc agtgtcctgc aggctcaaac cccaatgcga tttatcaggt gcttggatcc
     1921 ggtggaagta tggctagcat gactggcggg cagcaaatgg gcgggtctag cgcaatggca
     1981 tctatgaccg gaggtcaaca gatgggaggt tcctcagcca tggcgagtat gacaggtggc
     2041 caacagatgg gttaagaatt cgccagcaca gtggtcgacc aattccggtt attttccacc
     2101 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2161 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2221 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2281 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2341 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2401 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2461 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     2521 ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     2581 gataatacca tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     2641 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     2701 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     2761 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     2821 cttgacattg gggaatttag cgagagcctg acctattgca tctcccgccg tgcacagggt
     2881 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     2941 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     3001 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     3061 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     3121 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     3181 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     3241 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     3301 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     3361 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     3421 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     3481 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     3541 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     3601 agggcaaagg aatagagtag atgccgaccg ggatctatcg ataaaataaa agattttatt
     3661 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     3721 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc
     3781 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     3841 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     3901 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     3961 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     4021 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     4081 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     4141 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     4201 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     4261 cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt
     4321 acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4381 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4441 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4501 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4561 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4621 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4681 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4741 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4801 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4861 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4921 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4981 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5041 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5101 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5161 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5221 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc
     5281 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5341 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5401 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5461 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5521 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5581 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5641 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5701 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5761 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5821 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5881 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5941 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6001 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6061 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6121 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6181 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6241 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6301 aaagtgccac ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     6361 ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     6421 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag
     6481 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     6541 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     6601 tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat
     6661 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     6721 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     6781 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
     6841 caccgggagg caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
     6901 actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc
     6961 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
     7021 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
     7081 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     7141 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
     7201 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
     7261 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
     7321 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
     7381 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
     7441 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
     7501 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
     7561 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
     7621 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca
     7681 gtgtggtggt acgtaggaat tcccacc
//
