LOCUS Exported 8190 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLV.PGK.mLmx1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8190) AUTHORS Pfisterer U, Kirkeby A, Torper O, Wood J, Nelander J, Dufour A, Bjorklund A, Lindvall O, Jakobsson J, Parmar M TITLE Direct conversion of human fibroblasts to dopaminergic neurons. JOURNAL Proc Natl Acad Sci U S A. 2011 Jun 21;108(25):10343-8. Epub 2011 Jun 6. PUBMED 21646515 REFERENCE 2 (bases 1 to 8190) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8190 /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" RBS 5204..5215 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" primer_bind complement(6353..6373) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6771..6782) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6931..7164 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7236..7357 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7273..7292) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7327..7346 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 7376..7511 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7438..7457 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(7543..7562) /label=T7 /note="T7 promoter, forward primer" promoter complement(7544..7562) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7572..7589) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7572..7588) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7581..7603) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7733..8188 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7820..7839) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8030..8051 /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 atccatgttg gacggcctga 5161 agatggagga gaactttcaa agtgcgattg agacctcggc atctttctcc tctttgctgg 5221 gcagagcggt gagccccaag tctgtctgcg agggctgtca gcgggtcatc tcggacaggt 5281 ttctgctgcg gctcaacgac agcttctggc acgagcaatg cgtgcagtgt gcctcctgca 5341 aagagcccct ggagaccacc tgcttctacc gggacaagaa gctctactgc aagtaccact 5401 acgagaaact gtttgctgtc aaatgtgggg gctgcttcga ggccattgcg cccaatgagt 5461 ttgtcatgcg tgcccagaag agcgtatacc acctgagctg cttctgctgc tgcgtctgtg 5521 agcgacagct gcagaagggt gacgagtttg tcctgaagga gggccagctg ctctgcaaag 5581 gggactatga gaaagaacgg gagctgctga gcctggtgag ccctgcggcc tcagactcag 5641 gcaaaagcga tgatgaggag agcctttgca agtcagccca tggggcagga aaaggagcat 5701 cagaggacgg caaggaccat aagcgaccca aacgtcccag aaccatcctg accactcagc 5761 agaggagagc attcaaggcc tcgtttgaag tatcctccaa gccctgcaga aaggtgaggg 5821 agactctggc tgcggagaca gggctgagtg tccgtgtggt tcaggtgtgg ttccagaacc 5881 agcgagccaa gatgaagaag ctggcccggc gacagcagca acagcaacag gaccaacaga 5941 acacccagag gctgacttct gctcagacaa atggtagtgg gaatgcgggc atggaaggga 6001 tcatgaaccc ctatacaacg ttgcccaccc cacagcagct gctggccatt gaacagagcg 6061 tctacaactc tgatcccttc cgacagggtc tcaccccacc ccagatgcct ggagatcaca 6121 tgcaccccta tggtgctgaa cctcttttcc atgacttgga tagtgatgac acatctctca 6181 gtaacctggg agactgcttc ctggcaacct cagaagctgg gcccctgcag tccagagtgg 6241 gaaaccccat tgaccatctg tactccatgc agaattccta tttcacctct tgagtcgaca 6301 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 6361 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 6421 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 6481 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 6541 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6601 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6661 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 6721 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6781 atccagcgga ccttccttcc cgcggcctgc tgccggctct agagcctctt ccgcgtcttc 6841 gccttcccgg gtcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc 6901 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac 6961 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc 7021 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 7081 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 7141 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat 7201 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg 7261 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 7321 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc 7381 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 7441 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 7501 ttttggaggc ctaggctttt gcgtcgagac gtacccaatt cgccctatag tgagtcgtat 7561 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 7621 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 7681 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc 7741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 7801 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 7861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 7921 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 7981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 8041 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 8101 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 8161 tttaacaaaa tattaacgtt tacaatttcc //