LOCUS       sequence-235535-        8475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of MyoD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8475)
  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 8475)
  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.MyoD.WPRE
FEATURES             Location/Qualifiers
     source          1..8475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    132..153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(168..173,174..191,192..198)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    206..222
                     /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     211..233
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     230..246
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     230..246
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     265..285
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        267..285
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        313..539
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             540..720
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    767..892
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1385..1618
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1803..1847
                     /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    2114..2231
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2277..2787
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2706..2724
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    4631..5219
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4684..4704)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5102..5113)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5306..5539
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5611..5732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5648..5667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5702..5721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5772..5907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5927..5946)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5928..5946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5956..5973)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5956..5972)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5965..5987)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6114..6569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6201..6220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6411..6432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6595..6699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6700..6768,6769..7560)
                     /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(6918..6937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7731..8319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8220..8239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(8473..8475,1..15)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
       61 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
      121 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
      181 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
      241 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
      301 ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga
      361 gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa
      421 ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa
      481 ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg
      541 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      601 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
      661 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
      721 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
      781 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
      841 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
      901 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
      961 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     1021 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     1081 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     1141 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     1201 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     1261 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1321 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1381 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1441 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1501 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1561 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1621 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1681 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     1741 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     1801 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     1861 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     1921 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     1981 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2041 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     2101 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     2161 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     2221 ttcaaaattt tatcgatcac gagactagcc tcgagaagct tgatatcgaa ttccacgggg
     2281 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
     2341 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
     2401 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
     2461 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
     2521 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
     2581 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
     2641 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
     2701 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
     2761 gaccgaatca ccgacctctc tccccagggg gatcccagtg tggtggtacg taggaattcg
     2821 gacacctctg acaggacagg acagggagga ggggtagagg acagccggtg tgcattccaa
     2881 cccacagaac ctttgtcatt gtactgttgg ggttccggag tggcagaaag ttaagacgac
     2941 tctcacggct tgggttgagg ctggacccag gaactgggat atggagcttc tatcgccgcc
     3001 actccgggac atagacttga caggccccga cggctctctc tgctcctttg agacagcaga
     3061 cgacttctat gatgatccgt gtttcgactc accagacctg cgcttttttg aggacctgga
     3121 cccgcgcctg gtgcacgtgg gagccctcct gaaaccggag gagcacgcac acttctctac
     3181 tgcggtgcac ccaggcccag gcgctcgtga ggatgagcat gtgcgcgcgc ccagcgggca
     3241 ccaccaggcg ggtcgctgct tgctgtgggc ctgcaaggcg tgcaagcgca agaccaccaa
     3301 cgctgatcgc cgcaaggccg ccaccatgcg cgagcgccgc cgcctgagca aagtgaatga
     3361 ggccttcgag acgctcaagc gctgcacgtc cagcaacccg aaccagcggc tacccaaggt
     3421 ggagatcctg cgcaacgcca tccgctacat cgaaggtctg caggctctgc tgcgcgacca
     3481 ggacgccgcg ccccctggcg ccgctgcctt ctacgcacct ggaccgctgc ccccaggccg
     3541 tggcagcgag cactacagtg gcgactcaga cgcgtccagc ccgcgctcca actgctctga
     3601 tggcatgatg gattacagcg gccccccaag cggcccccgg cggcagaatg gctacgacac
     3661 cgcctactac agtgaggcgg tgcgcgagtc caggccaggg aagagtgcgg ctgtgtcgag
     3721 cctcgactgc ctgtccagca tagtggagcg catctccaca gacagccccg ctgcgcctgc
     3781 gctgcttttg gcagatgcac caccagagtc gcctccgggt ccgccagagg gggcatccct
     3841 aagcgacaca gaacagggaa cccagacccc gtctcccgac gccgcccctc agtgtcctgc
     3901 aggctcaaac cccaatgcga tttatcaggt gctttgagag atcgactgca gcagcagagg
     3961 gcgcaccacc gtaggcactc ctggggatgg tgcccctggt tcttcacgcc caaaagatga
     4021 agcttaaatg acactcttcc caactgtcct ttcgaagccg ttcttccaga gggaagggaa
     4081 gagcagaagt ctgtcctaga tccagcccca aagaaaggac atagtccttt ttgttgttgt
     4141 tgttgtagtc cttcagttgt ttgtttgttt tttcatgcgg ctcacagcga aggccacttg
     4201 cactctggct gcacctcact gggccagagc tgatccttga gtggccaggc gctcttcctt
     4261 tcctcatagc acaggggtga gccttgcaca cctaagccct gccctccaca tccttttgtt
     4321 tgtcactttc tggagccctc ctggcaccca cttttcccca cagcttgcgg aggccactca
     4381 ggtctcaggt gtaacaggtg taaccatacc ccactctccc ccttcccgcg gttcaggacc
     4441 acttattttt ttatataaga cttttgtaat ctattcgtgt aaataagagt tgcttggcca
     4501 gagcgggagc cccttgggct atatttatct cccaggcatg ctgtgtagtg caacaaaaac
     4561 tttgtatgtt tattcctcaa gcgggcgagt caggtgttgg aaatccgaat tcgccagcac
     4621 agtggtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4681 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     4741 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     4801 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     4861 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     4921 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     4981 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5041 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5101 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5161 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5221 tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5281 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5341 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5521 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     5581 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5701 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5761 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5821 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5881 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     5941 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6001 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6061 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6121 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6181 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6241 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6301 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6361 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6421 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6481 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6541 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     6601 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6661 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6721 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6781 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6841 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6901 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6961 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7021 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7081 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7141 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7201 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7261 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7321 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7381 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7441 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7501 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7561 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7621 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7681 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7741 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7801 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7861 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7921 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7981 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8041 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8101 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8161 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8221 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8281 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8341 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8401 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8461 acgaccgagc gcagc
//
