LOCUS Exported 8421 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLV.PGK.mFoxa2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8421) 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 8421) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8421 /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" primer_bind complement(6584..6604) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7002..7013) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7162..7395 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7467..7588 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7504..7523) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7558..7577 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 7607..7742 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7669..7688 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(7774..7793) /label=T7 /note="T7 promoter, forward primer" promoter complement(7775..7793) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7803..7820) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7803..7819) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7812..7834) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7964..8419 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8051..8070) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8261..8282 /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 atccatgctg ggagccgtga 5161 agatggaagg gcacgagcca tccgactgga gcagctacta cgcggagccc gagggctact 5221 cttccgtgag caacatgaac gccggcctgg ggatgaatgg catgaacaca tacatgagca 5281 tgtccgcggc tgccatgggc ggcggttccg gcaacatgag cgcgggctcc atgaacatgt 5341 catcctatgt gggcgctgga atgagcccgt cgctagctgg catgtccccg ggcgccggcg 5401 ccatggcggg catgagcggc tcagccgggg cggccggcgt ggcgggcatg ggacctcacc 5461 tgagtccgag tctgagcccg ctcgggggac aggcggccgg ggccatgggt ggccttgccc 5521 cctacgccaa catgaactcg atgagcccca tgtacgggca ggccggcctg agccgcgctc 5581 gggaccccaa gacataccga cgcagctaca cacacgccaa acctccctac tcgtacatct 5641 cgctcatcac catggccatc cagcagagcc ccaacaagat gctgacgctg agcgagatct 5701 atcagtggat catggacctc ttccctttct accggcagaa ccagcagcgc tggcagaact 5761 ccatccgcca ctctctctcc ttcaacgact gctttctcaa ggtgccccgc tcgccagaca 5821 agcctggcaa gggctccttc tggaccctgc acccagactc gggcaacatg ttcgagaacg 5881 gctgctacct gcgccgccag aagcgcttca agtgtgagaa gcaactggca ctgaaggaag 5941 ccgcgggtgc ggccagtagc ggaggcaaga agaccgctcc tgggtcccag gcctctcagg 6001 ctcagctcgg ggaggccgcg ggctcggcct ccgagactcc ggcgggcacc gagtcccccc 6061 attccagcgc ttctccgtgt caggagcaca agcgaggtgg cctaagcgag ctaaagggag 6121 cacctgcctc tgcgctgagt cctcccgagc cggcgccctc gcctgggcag cagcagcagg 6181 ctgcagccca cctgctgggc ccacctcacc acccaggcct gccaccagag gcccacctga 6241 agcccgagca ccattacgcc ttcaaccacc ccttctctat caacaacctc atgtcgtccg 6301 agcagcaaca tcaccacagc caccaccacc atcagcccca caaaatggac ctcaaggcct 6361 acgaacaggt catgcactac ccagggggct atggttcccc catgccaggc agcttggcca 6421 tgggcccagt cacgaacaaa gcgggcctgg atgcctcgcc cctggctgca gacacttcct 6481 actaccaagg agtgtactcc aggcctatta tgaactcatc ctaagtcgac aatcaacctc 6541 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc 6601 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca 6661 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg 6721 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca 6781 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg 6841 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg 6901 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg 6961 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg 7021 accttccttc ccgcggcctg ctgccggctc tagagcctct tccgcgtctt cgccttcccg 7081 ggtcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 7141 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc 7201 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct 7261 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt 7321 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt 7381 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa 7441 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat 7501 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 7561 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac 7621 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 7681 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 7741 cctaggcttt tgcgtcgaga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc 7801 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 7861 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 7921 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc 7981 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 8041 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 8101 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 8161 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 8221 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact 8281 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 8341 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 8401 atattaacgt ttacaatttc c //