LOCUS Exported 8457 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Induce human iN conversion from fibroblast. ACCESSION . VERSION . KEYWORDS 1000.pCCLsin.cPPT.hPGK.Brn2.WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8457) AUTHORS Lau S, Rylander Ottosson D, Jakobsson J, Parmar M TITLE Direct neural conversion from human fibroblasts using self-regulating and nonintegrating viral vectors. JOURNAL Cell Rep. 2014 Dec 11;9(5):1673-80. doi: 10.1016/j.celrep.2014.11.017. Epub 2014 Dec 4. PUBMED 25482564 REFERENCE 2 (bases 1 to 8457) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8457 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 27..131 /gene="bla" /label=AmpR promoter CDS 132..992 /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(350..369) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1163..1751 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1652..1671 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1905..1922 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2039..2060 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2075..2105 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2113..2129 /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 2118..2140 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2137..2153 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2137..2153 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2172..2192 /label=T3 /note="T3 promoter, forward primer" promoter 2174..2192 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" enhancer 2293..2672 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2673..2876 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2827..2847 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 2891..3071 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 3115..3240 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3733..3966 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 4151..4195 /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 4462..4579 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 4626..5136 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 5055..5073 /label=hPGK-F /note="Human PGK promoter, forward primer" regulatory 5155..5164 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(6599..6619) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7017..7028) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7177..7410 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7482..7603 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7519..7538) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7573..7592 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 7643..7778 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7705..7724 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(7810..7829) /label=T7 /note="T7 promoter, forward primer" promoter complement(7811..7829) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7839..7856) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7839..7855) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7848..7870) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8000..8455 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8087..8106) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8297..8318 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttggc 2221 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat 2281 taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat 2341 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg 2401 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa 2461 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact 2521 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta 2581 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt 2641 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg 2701 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 2761 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc 2821 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt 2881 tagtgaaccg gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 2941 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 3001 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 3061 aatctctagc agtggcgccc gaacagggac ctgaaagcga aagggaaacc agagctctct 3121 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg 3181 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc 3241 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga 3301 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca 3361 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa 3421 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc 3481 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag 3541 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg 3601 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat 3661 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag 3721 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg 3781 cgcagcctca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca 3841 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg 3901 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca 3961 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa 4021 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg 4081 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa 4141 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa 4201 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg 4261 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg 4321 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga 4381 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg 4441 atctcgacgg tatcggttaa cttttaaaag aaaagggggg attggggggt acagtgcagg 4501 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat 4561 tacaaaaatt caaaatttta tcgatcacga gactagcctc gagaagcttg atatcgaatt 4621 cccacggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc 4681 tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc 4741 acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg 4801 cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga 4861 caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg 4921 gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcggg gcgcgccgag 4981 agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg 5041 ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc 5101 tccctcgttg accgaatcac cgacctctct ccccaggggg atccaccggt gctagccacc 5161 atggcgaccg cagcgtctaa ccactacagc ctgctcacct ccagcgcctc catcgtacat 5221 gccgagccgc ctggcggcat gcagcagggc gcagggggct accgcgaggc gcagagcctg 5281 gtgcagggcg actacggcgc gctgcagagc aacgggcacc cgctcagcca cgctcaccag 5341 tggatcaccg cgctgtccca cggcggcggc ggcgggggcg gcggcggcgg tggaggaggc 5401 gggggaggcg gcgggggagg cggcgacggc tccccgtggt ccaccagccc cctaggccag 5461 ccggacatca agccctcggt ggtggtacag cagggtggcc gaggcgacga gctgcacggg 5521 ccaggagcgc tgcagcaaca gcatcaacag caacagcaac agcagcagca gcagcagcag 5581 cagcagcagc agcaacagca gcagcaacaa cagcgaccgc cacatctggt gcaccacgct 5641 gccaaccacc atcccgggcc cggggcatgg cggagtgcgg cggctgcagc tcacctccct 5701 ccctccatgg gagcttccaa cggcggtttg ctctattcgc agccgagctt cacggtgaac 5761 ggcatgctgg gcgcaggagg gcagccggct gggctgcacc accacggcct gagggacgcc 5821 cacgatgagc cacaccatgc agaccaccac ccgcatccgc actctcaccc acaccagcaa 5881 ccgcccccgc cacctccccc acaaggccca ccgggccacc caggcgcgca ccacgacccg 5941 cactcggacg aggacacgcc gacctcagac gacctggagc agttcgccaa gcaattcaag 6001 cagaggcgga tcaaactcgg atttactcaa gcagacgtgg ggctggcgct tggcaccctg 6061 tacggcaacg tgttctcgca gaccaccatc tgcaggtttg aggccctgca gctgagcttc 6121 aagaacatgt gcaagctgaa gcctttgttg aacaagtggt tggaagaggc agactcatcc 6181 tcgggcagcc ccaccagcat agacaagatc gcagcgcaag ggcgcaaacg gaaaaagcgg 6241 acctccatcg aggtgagcgt caagggggct ctggagagcc atttcctcaa atgccctaag 6301 ccctcggccc aggagatcac ctccctcgcg gacagcttac agctggagaa ggaggtggtg 6361 agagtttggt tttgtaacag gagacagaaa gagaaaagga tgacccctcc cggagggact 6421 ctgccgggcg ccgaggatgt gtatgggggt agtagggaca cgccaccaca ccacggggtg 6481 cagacgcccg tccagtgagc taggaataag ggcgaattcg atatcaagct tatcgataag 6541 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 6601 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 6661 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 6721 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 6781 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 6841 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 6901 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 6961 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 7021 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctagag cctcttccgc 7081 gtcttcgcct tcccgggtcg agctcggtac ctttaagacc aatgacttac aaggcagctg 7141 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac 7201 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct 7261 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag 7321 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac 7381 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt 7441 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt 7501 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 7561 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc 7621 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 7681 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga 7741 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgcg tcgagacgta 7801 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt 7861 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc 7921 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc 7981 ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 8041 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 8101 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 8161 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 8221 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 8281 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 8341 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 8401 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcc //