LOCUS       sequence-235399-        7665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of a MyoD chimeric protein substituted with 
            the NeuroD2 bHLH domain and point mutations to prevent MyoD 
            interaction with PBX/MEIS: W96A, C98A, and S253P.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  AUTHORS   Fong AP, Yao Z, Zhong JW, Johnson NM, Farr GH 3rd, Maves L, Tapscott
            SJ
  TITLE     Conversion of MyoD to a neurogenic factor: binding site specificity 
            determines lineage.
  JOURNAL   Cell Rep. 2015 Mar 31;10(12):1937-46. doi: 
            10.1016/j.celrep.2015.02.055. Epub 2015 Mar 19.
  PUBMED    25801030
REFERENCE   2  (bases 1 to 7665)
  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.celrep.2015.02.055"; journalName: "Cell Rep"; date: 
            "2015-03-31- 31"; volume: "10"; issue: "12"; pages: "1937-46"
COMMENT     Sequence Label: pRRLSIN.cPPT.PGK.MD(ND2bHLH)WCS.WPRE
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      26..35
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1034..1622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1087..1107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1505..1516)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1709..1942
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2014..2135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2051..2070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2105..2124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2175..2310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(2331..2349)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2359..2376)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2359..2375)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2368..2390)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2517..2972
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2604..2623)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2814..2835
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2998..3102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3103..3171,3172..3963)
                     /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     complement(3321..3340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4134..4722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4623..4642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4876..4893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5010..5031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5046..5051,5052..5069,5070..5076)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5084..5100
                     /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)."
     primer_bind     5089..5111
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5108..5124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5108..5124
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5143..5163
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5145..5163
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5191..5417
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5418..5598
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5645..5770
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6263..6496
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6681..6725
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6992..7109
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7155..7665
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7584..7602
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 ggggatccca tcgattcgaa ttcccgccac catggagctt ctatcgccgc cactccggga
       61 catagacttg acaggccccg acggctctct ctgctccttt gagacagcag acgacttcta
      121 tgatgacccg tgtttcgact caccagacct gcgctttttt gaggacctgg acccgcgcct
      181 ggtgcacatg ggagccctcc tgaaaccgga ggagcacgca cacttcccta ctgcggtgca
      241 cccaggccca ggcgctcgtg aggatgagca tgtgcgcgcg cccagcgggc accaccaggc
      301 gggtcgctgc ttgctggcgg ccgccaaggc gtgcaagcgc aagaccacca acgctgatcg
      361 gcgccagaag gccaacgcgc gggagcgcaa ccgcatgcac gacctgaacg cggctctgga
      421 caacctgcgc aaggtggtgc cctgctactc caagacgcag aagctgtcca agatcgagac
      481 cctgcgcctg gccaagaact acatctgggc tctctcggag atcttgcgct ccgggcagga
      541 cgccgcgccc cctggcgccg ctgccttcta cgcacctgga ccgctgcccc caggccgtgg
      601 cagcgagcac tacagtggcg actcagatgc atccagcccg cgctccaact gctctgatgg
      661 catgatggat tacagcggcc ccccaagcgg cccccggcgg cagaatggct acgacaccgc
      721 ctactacagt gaggcggcgc gcgagtccag gccagggaag agtgcggctg tgtcgagcct
      781 cgactgcctg tcacccatag tggagcgcat ctccacagac agccccgctg cgcctgcgct
      841 gcttttggca gatgcaccac cagagtcgcc tccgggtccg ccagaggggg catccctaag
      901 cgacacagaa cagggaaccc agaccccgtc tcccgacgcc gcccctcagt gtcctgcagg
      961 ctcaaacccc aatgcgattt atcaggtgct ttgagtcgag cctctagaac tatagtgagt
     1021 cgtattactc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1081 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1141 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     1201 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     1261 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     1321 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     1381 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     1441 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     1501 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     1561 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     1621 gcctggaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     1681 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     1741 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     1801 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     1861 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     1921 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     1981 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     2041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     2161 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2221 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2281 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     2341 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2401 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2461 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     2521 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2581 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2641 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2701 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2761 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2821 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2881 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2941 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     3001 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3061 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3121 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3181 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3241 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3301 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3361 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3421 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3481 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3541 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3601 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3661 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3721 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3781 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3841 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3901 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3961 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4021 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4081 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4141 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4201 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4261 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4321 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4381 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4441 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4501 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4561 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4621 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4681 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4741 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4801 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4861 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4921 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4981 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     5041 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     5101 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     5161 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     5221 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     5281 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     5341 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     5401 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5461 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5521 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5581 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     5641 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     5701 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     5761 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     5821 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     5881 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     5941 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     6001 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     6061 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     6121 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     6181 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     6241 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     6301 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     6361 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     6421 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     6481 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     6541 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     6601 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     6661 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     6721 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     6781 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     6841 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg
     6901 acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat
     6961 tagtgaacgg atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt
     7021 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     7081 aaaaacaaat tacaaaaatt caaaatttta tcgatcacga gactagcctc gagaagcttg
     7141 atatcgaatt ccacggggtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag
     7201 ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc
     7261 acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc
     7321 ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc
     7381 cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag
     7441 ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg
     7501 cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg
     7561 tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg
     7621 gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccag
//
