LOCUS       sequence-235538-        7805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of NeuroD2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7805)
  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 7805)
  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.NeuroD2.WPRE
FEATURES             Location/Qualifiers
     source          1..7805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1095..1683
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1148..1168)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1566..1577)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1770..2003
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2075..2196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2112..2131)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2166..2185
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2236..2371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2391..2410)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2392..2410)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2420..2437)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2420..2436)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2429..2451)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2578..3033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2665..2684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2875..2896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3059..3163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3164..3232,3233..4024)
                     /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(3382..3401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4195..4783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4684..4703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4937..4954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5071..5092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5107..5112,5113..5130,5131..5137)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5145..5161
                     /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     5150..5172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5169..5185
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5169..5185
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5204..5224
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5206..5224
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5252..5478
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5479..5659
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5706..5831
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6324..6557
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6742..6786
                     /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    7053..7170
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7216..7726
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7645..7663
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 cgacggcgac gacgacgagc cgaggagcga caagggcgac gcgccgccgc agcctcctcc
       61 tgctcccggg tcgggggctc caggacccgc ccgggccgcc aagccagtgt ctcttcgtgg
      121 aggagaagag atccctgaac ccacgttggc tgaggtcaag gaggaaggag agctgggcgg
      181 cgaggaggag gaggaagagg aggaggagga aggactggac gaggcggaag gcgagcggcc
      241 caagaagcgc gggccgaaga aacgcaagat gaccaaggcg cgtctggagc gctccaagct
      301 gcggcgacag aaggccaacg cgcgggagcg caaccgcatg cacgacctga acgcggctct
      361 ggacaacctg cgcaaggtgg tgccctgcta ctccaagacg cagaagctgt ccaagatcga
      421 gaccctgcgc ctggccaaga actacatctg ggctctctcg gagatcttgc gctccgggaa
      481 gcggccggat ctggtgtcct acgtgcagac tctgtgcaag gggctgtcac agcccaccac
      541 gaatctggtg gccggctgcc tgcagctaaa ctctcgtaac ttcctcacgg agcagggcgc
      601 ggacggcgcc ggccgctttc acggctcggg tggcccgttc gccatgcatc cgtacccata
      661 cccgtgctcc cgcctggcag gcgcacagtg tcaggcggct ggcggcctgg gcggaggcgc
      721 ggcgcacgcc ctgcggaccc acggctactg cgccgcctac gagacgctgt acgcggcggc
      781 cggtggcggc ggcgctagcc cggactacaa cagctccgag tacgagggtc cactcagtcc
      841 cccgctctgt ctcaacggca acttctcgct caagcaggac tcgtcccccg atcacgagaa
      901 gagctaccac tactctatgc actactcggc gctgcccggc tcacggccca cgggccacgg
      961 gctggtcttc ggctcgtcgg ccgtgcgcgg gggcgtccac tccgagaatc tcttgtctta
     1021 cgatatgcac cttcaccacg atcggggccc catgtacgag gagctcaacg catttttcca
     1081 taactgaggt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1141 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1201 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     1261 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     1321 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     1381 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1441 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     1501 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     1561 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     1621 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     1681 cgcctggaat tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     1741 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     1801 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     1861 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1921 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1981 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     2041 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     2101 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2161 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     2221 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2281 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2341 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     2401 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2461 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     2521 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     2581 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2641 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2701 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2761 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2821 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2881 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2941 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3001 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     3061 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3121 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3541 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     3601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3721 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4021 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4081 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4141 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4321 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4861 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4921 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4981 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5041 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5101 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5161 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     5221 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     5281 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     5341 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     5401 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     5461 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5521 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5581 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5641 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     5701 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     5761 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     5821 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     5881 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     5941 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     6001 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     6061 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     6121 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     6181 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     6241 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     6301 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     6361 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     6421 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     6481 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     6541 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     6601 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     6661 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     6721 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     6781 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     6841 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     6901 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     6961 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     7021 ttagtgaacg gatctcgacg gtatcggtta acttttaaaa gaaaaggggg gattgggggg
     7081 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     7141 caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg agactagcct cgagaagctt
     7201 gatatcgaat tccacggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca
     7261 gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg
     7321 cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc
     7381 cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg
     7441 ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca
     7501 gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg
     7561 gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt
     7621 gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc
     7681 ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg atccgccacc
     7741 atgctgaccc gcctgttcag cgagcccggc ctcctctcgg acgtgcccaa gttcgccagc
     7801 tgggg
//
