LOCUS       sequence-322392-        9129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9129)
  AUTHORS   Yoo AS, Sun AX, Li L, Shcheglovitov A, Portmann T, Li Y, Lee-Messer 
            C, Dolmetsch RE, Tsien RW, Crabtree GR
  TITLE     MicroRNA-mediated conversion of human fibroblasts to neurons.
  JOURNAL   Nature. 2011 Jul 13. doi: 10.1038/nature10323.
  PUBMED    21753754
REFERENCE   2  (bases 1 to 9129)
  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"; journalName: "Nature. 
            2011 Jul 13. doi: 10.1038/nature10323."
COMMENT     Sequence Label: phASCL1-N106
FEATURES             Location/Qualifiers
     source          1..9129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..65
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             140..538
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    562..1150
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(615..635)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1033..1044)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1357..1990
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(2118..2134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2118..2134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2131..2153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2142..2158
                     /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(2166..2172,2173..2190,2191..2196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2211..2232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2349..2366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2520..3108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2600..2619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3279..4070,4071..4139))
                     /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     3902..3921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4140..4244)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    4292..4426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4329..4348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4383..4402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4594..5227
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5274..5399
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5896..6129
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6314..6358
                     /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    6621..6736
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        6814..7025
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          7026..7964
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7921..7941
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7986..7995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        join(8743..9129,1..113)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8778..8801)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
       61 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
      121 gcccaagctt cctaggacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
      181 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
      241 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
      301 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
      361 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
      421 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
      481 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaacc
      541 cgggcggggc gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact
      601 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
      661 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
      721 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
      781 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
      841 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
      901 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
      961 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     1021 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     1081 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     1141 gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa
     1201 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     1261 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     1321 tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac
     1381 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc
     1441 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc
     1501 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt
     1561 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt
     1621 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga
     1681 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg
     1741 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt
     1801 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     1861 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     1921 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     1981 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     2041 tgaatatcag agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag
     2101 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2161 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2221 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2281 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2341 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2401 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2461 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2521 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2581 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2641 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2701 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2761 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2821 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2881 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2941 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3001 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3061 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3121 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3181 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3241 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3301 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3361 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3421 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3481 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3541 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3601 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3661 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3721 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3781 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3841 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3901 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3961 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4021 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4081 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4141 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4201 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4261 gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta
     4321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4381 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca
     4441 agctaaccaa aatcatccca aacttcccac cccataccct attaccactg ccaattacct
     4501 gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg
     4561 tatttgttaa atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa
     4621 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
     4681 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
     4741 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
     4801 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
     4861 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
     4921 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
     4981 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     5041 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5101 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5161 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5221 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     5281 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     5341 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     5401 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     5461 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     5521 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     5581 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     5641 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     5701 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc
     5761 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     5821 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     5881 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     5941 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     6001 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     6061 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     6121 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     6181 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     6241 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     6301 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     6361 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     6421 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     6481 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     6541 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     6601 cggatctcga cggtatcgcc tttaaaagaa aaggggggat tggggggtac agtgcagggg
     6661 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     6721 caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt tatcgatgag
     6781 gccctttcgt cttcactcga ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     6841 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
     6901 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     6961 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     7021 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
     7081 gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg
     7141 ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt
     7201 gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg
     7261 cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga
     7321 cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt
     7381 cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga
     7441 ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc
     7501 ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg
     7561 cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga
     7621 gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga
     7681 aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt
     7741 ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg
     7801 ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt
     7861 ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc
     7921 tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaggatccg
     7981 cggccgccac catggaaagc tctgccaaga tggagagcgg cggcgccggc cagcagcccc
     8041 agccgcagcc ccagcagccc ttcctgccgc ccgcagcctg tttctttgcc acggccgcag
     8101 ccgcggcggc cgcagccgcc gcagcggcag cgcagagcgc gcagcagcag cagcagcagc
     8161 agcagcagca gcagcaggcg ccgcagctga gaccggcggc cgacggccag ccctcagggg
     8221 gcggtcacaa gtcagcgccc aagcaagtca agcgacagcg ctcgtcttcg cccgaactga
     8281 tgcgctgcaa acgccggctc aacttcagcg gctttggcta cagcctgccg cagcagcagc
     8341 cggccgccgt ggcgcgccgc aacgagcgcg agcgcaaccg cgtcaagttg gtcaacctgg
     8401 gctttgccac ccttcgggag cacgtcccca acggcgcggc caacaagaag atgagtaagg
     8461 tggagacact gcgctcggcg gtcgagtaca tccgcgcgct gcagcagctg ctggacgagc
     8521 atgacgcggt gagcgccgcc ttccaggcag gcgtcctgtc gcccaccatc tcccccaact
     8581 actccaacga cttgaactcc atggccggct cgccggtctc atcctactcg tcggacgagg
     8641 gctcttacga cccgctcagc cccgaggagc aggagcttct cgacttcacc aactggttct
     8701 gaggccgcgc cggctctaga tcgcgaacgc gtgaattcta ccgggtaggg gaggcgcttt
     8761 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     8821 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     8881 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     8941 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
     9001 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
     9061 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
     9121 tcaggggcg
//
