LOCUS Exported 6956 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Destination vector for TALE-VP64 construction. ACCESSION . VERSION . KEYWORDS pTALE-STARVP64-T SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6956) 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 6956) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6956 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 383..658 /label=chicken beta-actin promoter intron 659..1667 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1591..1613 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" regulatory 1731..1740 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1740..1805 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 1812..1832 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2615..2635 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2660..2809 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 2831..2884 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" primer_bind 3379..3399 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(3563..3580) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3569..3793 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3840..3856) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3840..3856) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3853..3875) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3864..3880 /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(3888..3918) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3933..3954 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4013..4208 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4059..4194 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4121..4140 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4214..4348 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4416..4433) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4587..5175) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4667..4686) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5346..6206) /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 5969..5988 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 6809..6943 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc 421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg 481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg 541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc 601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg 661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg 721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg 781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct 841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt 901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg 961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt 1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg 1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct 1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg 1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg 1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc 1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac 1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag 1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt 1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg 1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct 1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg 1681 tgctggttgt tgtgctgtct catcattttg gcaaagaatt cctcgaggcc gccaccatgg 1741 actacaaaga ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg 1801 acaagatggc ccccaagaag aagaggaagg tgggcattca ccgcggggta cctatggtgg 1861 acttgaggac actcggttat tcgcaacagc aacaggagaa aatcaagcct aaggtcagga 1921 gcaccgtcgc gcaacaccac gaggcgcttg tggggcatgg cttcactcat gcgcatattg 1981 tcgcgctttc acagcaccct gcggcgcttg ggacggtggc tgtcaaatac caagatatga 2041 ttgcggccct gcccgaagcc acgcacgagg caattgtagg ggtcggtaaa cagtggtcgg 2101 gagcgcgagc acttgaggcg ctgctgactg tggcgggtga gcttaggggg cctccgctcc 2161 agctcgacac cgggcagctg ctgaagatcg cgaagagagg gggagtaaca gcggtagagg 2221 cagtgcacgc ctggcgcaat gcgctcaccg gggccccctt gaacctgcag gatacgttta 2281 tcagtgataa agtgtcaagc atgacaaagt tgcagccgaa tacagtggta tcctgcaggt 2341 gctctgtcag gaccacgggc ttacgccaca acaggtggtg gcgattgcat ctaatggagg 2401 cggacgccct gccttggagt ccatcgtagc ccaattgtcc aggcccgatc ccgcgttggc 2461 tgcgttaacg aatgaccatc tggtggcgtt ggcatgtctt ggtggacgac ccgcgctcga 2521 tgcagtcaaa aagggtctgc ctcatgctcc cgcattgatc aaaagaacca accggcggat 2581 tcccgagaga acttcccatc gagtcgcggc tagccccaag aagaagagaa aggtggaggc 2641 cagcggttcc ggacgggctg acgcattgga cgattttgat ctggatatgc tgggaagtga 2701 cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg actttgacct 2761 cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga ttaactctag 2821 aggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg 2881 cccaaccgag tacaagccca cggtgcgcct cgccacccgc gacgaacgtc cccagggccg 2941 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 3001 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 3061 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 3121 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 3181 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 3241 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 3301 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 3361 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 3421 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 3481 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga 3541 tgatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 3601 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 3661 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 3721 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 3781 gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc gtaatcatgg 3841 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 3901 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 3961 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca 4021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4141 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4201 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 4261 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 4321 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg 4381 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg 4441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccgcatac 4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat 4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt 5041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 5281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 5341 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 5401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 5461 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 5521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5641 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 5821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 5881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 5941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6181 gcgacacgga aatgttgaat actcatactc ttcctttttc aatgaccgct ttggaaaaac 6241 aaagactgta tttcctggaa attaatgttt attcaataaa ctgtgtattc agctatatca 6301 catagtggtg aggctgaaat gaggcgggaa gaggcggttg gggcttaatt atatcaattt 6361 gggtggcccc acagcgcctc caaggcgcca gtcctgtttt gacaagttgc ctctggaagc 6421 ctctacaatg cctctcttct ttttctccag agtaagcgga ggccaggggc ccccggcctc 6481 tgcttaatac taaaaaaaac agctgttgtc atagtaatga ttgggtggaa acattccagg 6541 cctgggtgga gaggcttttt gcttcctctt gcaaaaccac actgacattc caggcctggg 6601 tggagaggct ttttgcttcc tcttgcaaaa ccacactgcc ctctggaggg cagttgccta 6661 gcaactaact aaaagaggat gtcgcacggc cagctgcggt cagttagtca cttcctgctt 6721 aactgacttg acattttcta ttttaagagt cgggaggaaa attactgtgt tggaggccct 6781 ccgccatctt ctgaagctga atcgaattaa cttgtttatt gcagcttata atggttacaa 6841 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6901 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatatc atcgtc //