LOCUS Exported 6205 bp ds-DNA linear SYN 17-AUG-2021 DEFINITION Destination vector for TALE-VP160 construction. ACCESSION . VERSION . KEYWORDS pTALE-STARVP160-G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6205) AUTHORS Gogolok S, Garcia-Diaz C, Pollard SM TITLE STAR: a simple TAL effector assembly reaction using isothermal assembly. JOURNAL Sci Rep. 2016 Sep 12;6:33209. doi: 10.1038/srep33209. PUBMED 27615025 REFERENCE 2 (bases 1 to 6205) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6205 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 465..487 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 549..568 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 605..614 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 614..679 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 686..706 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1489..1509 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1552..1935 /codon_start=1 /product="10 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)" /label=VP160 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGS DALDDFDLDMLGSDALDDFDLDML" CDS 1957..2013 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 2014..2724 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(2159..2177) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2427..2446 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2822..2839) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2828..3052 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3099..3115) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3099..3115) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3112..3134) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3123..3139 /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(3147..3177) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3192..3213 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3272..3467 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3318..3453 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3380..3399 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3473..3607 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3675..3692) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3846..4434) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3926..3945) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4605..5465) /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 5228..5247 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 6058..6192 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg 61 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg 121 ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc 181 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag 241 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg 301 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg 361 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg 421 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg 481 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca 541 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattcctcga 601 ggccgccacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta 661 caaggatgac gatgacaaga tggcccccaa gaagaagagg aaggtgggca ttcaccgcgg 721 ggtacctatg gtggacttga ggacactcgg ttattcgcaa cagcaacagg agaaaatcaa 781 gcctaaggtc aggagcaccg tcgcgcaaca ccacgaggcg cttgtggggc atggcttcac 841 tcatgcgcat attgtcgcgc tttcacagca ccctgcggcg cttgggacgg tggctgtcaa 901 ataccaagat atgattgcgg ccctgcccga agccacgcac gaggcaattg taggggtcgg 961 taaacagtgg tcgggagcgc gagcacttga ggcgctgctg actgtggcgg gtgagcttag 1021 ggggcctccg ctccagctcg acaccgggca gctgctgaag atcgcgaaga gagggggagt 1081 aacagcggta gaggcagtgc acgcctggcg caatgcgctc accggggccc ccttgaacct 1141 gcaggatacg tttatcagtg ataaagtgtc aagcatgaca aagttgcagc cgaatacagt 1201 ggtatcctgc aggtgctctg tcaggaccac gggcttacgc cacaacaggt ggtggcgatt 1261 gcatctaatc acggcggacg ccctgccttg gtgtccatcg tagcccaatt gtccaggccc 1321 gatcccgcgt tggctgcgtt aacgaatgac catctggtgg cgttggcatg tcttggtgga 1381 cgacccgcgc tcgatgcagt caaaaagggt ctgcctcatg ctcccgcatt gatcaaaaga 1441 accaaccggc ggattcccga gagaacttcc catcgagtcg cggctagccc caagaagaag 1501 agaaaggtgg aggccagcgg ttccggacag tggagacggc gggctcgcgc cgacgcgctg 1561 gacgatttcg atctcgacat gctgggttct gatgccctcg atgactttga cctggatatg 1621 ttgggaagcg acgcattgga tgactttgat ctggacatgc tcggctccga tgctctggac 1681 gatttcgatc tcgatatgtt agggtcagac gcactggatg atttcgacct tgatatgttg 1741 ggaagcgatg cccttgatga tttcgacctg gacatgctcg gcagcgacgc cctggacgat 1801 ttcgatctgg acatgctggg gtccgatgcc ttggatgatt ttgacttgga tatgctgggg 1861 agtgatgccc tggacgactt tgacctggac atgctgggct ccgatgcgct cgatgacttc 1921 gatttggata tgttgattaa ctctagaggc agtggagcca ctaacttctc cctgttgaaa 1981 caagcagggg atgtcgaaga gaatcccggg ccaatggtga gcaagggcga ggaggataac 2041 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc 2101 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 2161 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag 2221 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag 2281 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg 2341 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg 2401 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag 2461 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg 2521 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag 2581 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac 2641 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc 2701 ggcatggacg agctgtacaa gtaacgcccg ccccacgacc cgcagcgccc gaccgaaagg 2761 agcgcacgac cccatgcatc gatgatctag agctccgtct caagctgagc tcgctgatca 2821 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 2881 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 2941 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 3001 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 3061 gcggaaagaa cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga 3121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 3181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 3241 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc 3301 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3361 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 3421 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt 3481 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 3541 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 3601 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 3661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 3721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 3781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 3841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 3901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 3961 gtgcgctctc ctgttccgac cctgccgctt accgcatacc tgtccgcctt tctcccttcg 4021 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 4081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 4141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 4201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 4261 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 4321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 4381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 4441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 4501 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 4561 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 4621 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 4681 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 4741 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 4801 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 4861 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 4921 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 4981 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 5041 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 5101 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 5161 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 5221 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 5281 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 5341 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 5401 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 5461 ctcatactct tcctttttca atgaccgctt tggaaaaaca aagactgtat ttcctggaaa 5521 ttaatgttta ttcaataaac tgtgtattca gctatattca catagtggtg aggctgaaat 5581 gaggcgggaa gaggcggttg gggcttaatt atatcaattt gggtggcccc acagcgcctc 5641 caaggcgcca gtcctgtttt gacaagttgc ctctggaagc ctctcttctt tttctccaga 5701 gtaagcggag gccaggggcc cccggcctct gcttaatact aaaaaaaaca gctgttgtca 5761 tagtaatgat tgggtggaaa cattccaggc ctgggtggag aggctttttg cttcctcttg 5821 caaaaccaca ctgacattcc aggcctgggt ggagaggctt tttgcttcct cttgcaaaac 5881 cacactgccc tctggagggc agttgcctag caactaatta aaagaggatg tcgcacggcc 5941 agctgcggtc agttagtcac ttcctgctta actgacttga cattttctat tttaagagtc 6001 gggaggaaaa ttactgtgtt ggaggccctc cgccatcttc tgaagctgaa tcgaattaac 6061 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 6121 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 6181 catgtctgga tctgatatca tcgtc //