LOCUS Exported 7976 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FUW-tetO-lox-hKLF4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7976) AUTHORS Frank Soldner, Dirk Hockemeyer, Caroline Beard, Qing Gao, George W. Bell, Elizabeth G. Cook, Gunnar Hargus, Alexandra Blak, Oliver Cooper, Maisam Mitalipova, Ole Isacson, Rudolf Jaenisch TITLE Parkinson's Disease Patient-Derived Induced Pluripotent Stem Cells Free of Viral Reprogramming Factors JOURNAL Cell, Volume 136, Issue 5, 964-977, 6 March 2009 REFERENCE 2 (bases 1 to 7976) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7976 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 7..386 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 388..586 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 541..561 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 604..784 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 831..956 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1449..1682 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1867..1911 /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 2209..2326 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2385..2655 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 2688..2725 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2722..2746 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 4365..4953 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4418..4438) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4836..4847) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4956..4972) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4957..4973) /label=pBluescriptKS /note="For pBluescript vector" protein_bind complement(5477..5510) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." LTR 5524..5704 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5730..5747) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5736..5847 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin complement(5993..6581) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6073..6092) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6752..7612) /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 7375..7394 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7613..7717) /gene="bla" /label=AmpR promoter primer_bind complement(7792..7811) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc 61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt 601 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 661 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 721 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 781 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac 841 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta 901 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta 961 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga 1021 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta 1081 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat 1141 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt 1201 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag 1261 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga 1321 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa 1381 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca 1441 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca 1501 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag 1561 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc 1621 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc 1681 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct 1741 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac 1801 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag 1861 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg 1921 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg 1981 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat 2041 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga 2101 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg 2161 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa 2221 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat 2281 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag 2341 ggacagcaga gatccagttt ggttaatgat ctcgagttta ccactcccta tcagtgatag 2401 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag 2461 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag 2521 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa 2581 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc 2641 tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgagta ggcgtgtacg 2701 gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca 2761 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccccga 2821 attccttctc ttcgttgact ttggggttca ggtgccccag ctgcttcggg ctgccgagga 2881 ccttctgggc ccccacatta atgaggcagc cacctggcga gtctgacatg gctgtcagcg 2941 acgcgctgct cccatctttc tccacgttcg cgtctggccc ggcgggaagg gagaagacac 3001 tgcgtcaagc aggtgccccg aataaccgct ggcgggagga gctctcccac atgaagcgac 3061 ttcccccagt gcttcccggc cgcccctatg acctggcggc ggcgaccgtg gccacagacc 3121 tggagagcgg cggagccggt gcggcttgcg gcggtagcaa cctggcgccc ctacctcgga 3181 gagagaccga ggagttcaac gatctcctgg acctggactt tattctctcc aattcgctga 3241 cccatcctcc ggagtcagtg gccgccaccg tgtcctcgtc agcgtcagcc tcctcttcgt 3301 cgtcgccgtc gagcagcggc cctgccagcg cgccctccac ctgcagcttc acctatccga 3361 tccgggccgg gaacgacccg ggcgtggcgc cgggcggcac gggcggaggc ctcctctatg 3421 gcagggagtc cgctccccct ccgacggctc ccttcaacct ggcggacatc aacgacgtga 3481 gcccctcggg cggcttcgtg gccgagctcc tgcggccaga attggacccg gtgtacattc 3541 cgccgcagca gccgcagccg ccaggtggcg ggctgatggg caagttcgtg ctgaaggcgt 3601 cgctgagcgc ccctggcagc gagtacggca gcccgtcggt catcagcgtc agcaaaggca 3661 gccctgacgg cagccacccg gtggtggtgg cgccctacaa cggcgggccg ccgcgcacgt 3721 gccccaagat caagcaggag gcggtctctt cgtgcaccca cttgggcgct ggaccccctc 3781 tcagcaatgg ccaccggccg gctgcacacg acttccccct ggggcggcag ctccccagca 3841 ggactacccc gaccctgggt cttgaggaag tgctgagcag cagggactgt caccctgccc 3901 tgccgcttcc tcccggcttc catccccacc cggggcccaa ttacccatcc ttcctgcccg 3961 atcagatgca gccgcaagtc ccgccgctcc attaccaaga gctcatgcca cccggttcct 4021 gcatgccaga ggagcccaag ccaaagaggg gaagacgatc gtggccccgg aaaaggaccg 4081 ccacccacac ttgtgattac gcgggctgcg gcaaaaccta cacaaagagt tcccatctca 4141 aggcacacct gcgaacccac acaggtgaga aaccttacca ctgtgactgg gacggctgtg 4201 gatggaaatt cgcccgctca gatgaactga ccaggcacta ccgtaaacac acggggcacc 4261 gcccgttcca gtgccaaaaa tgcgaccgag cattttccag gtcggaccac ctcgccttac 4321 acatgaagag gcatttttaa gaattcgata tcaagcttat cgataatcaa cctctggatt 4381 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 4441 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 4501 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 4561 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 4621 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 4681 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 4741 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct 4801 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 4861 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 4921 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag 4981 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc 5041 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac 5101 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg 5161 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac 5221 aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga 5281 cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg 5341 aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga 5401 gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc 5461 tgcatccgga ctcgagataa cttcgtataa tgtatgctat acgaagttat tccggactgt 5521 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 5581 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 5641 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 5701 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 5761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 5821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 5881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgtgag 5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 6421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 6601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 7741 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg 7801 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga 7861 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa 7921 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatata //