LOCUS Exported 7341 bp ds-DNA circular SYN 13-MAY-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 7341) 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 7341) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7341 /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 2678..3061 /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 3083..3139 /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 3140..3850 /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(3285..3303) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3553..3572 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(3948..3965) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3954..4178 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4225..4241) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4225..4241) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4238..4260) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4249..4265 /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(4273..4303) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4318..4339 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4398..4593 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4444..4579 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4506..4525 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4599..4733 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4801..4818) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4972..5560) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5052..5071) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5731..6591) /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 6354..6373 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 7194..7328 /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 ctaatcacgg 2401 cggacgccct gccttggtgt 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 ggacagtgga gacggcgggc tcgcgccgac gcgctggacg atttcgatct 2701 cgacatgctg ggttctgatg ccctcgatga ctttgacctg gatatgttgg gaagcgacgc 2761 attggatgac tttgatctgg acatgctcgg ctccgatgct ctggacgatt tcgatctcga 2821 tatgttaggg tcagacgcac tggatgattt cgaccttgat atgttgggaa gcgatgccct 2881 tgatgatttc gacctggaca tgctcggcag cgacgccctg gacgatttcg atctggacat 2941 gctggggtcc gatgccttgg atgattttga cttggatatg ctggggagtg atgccctgga 3001 cgactttgac ctggacatgc tgggctccga tgcgctcgat gacttcgatt tggatatgtt 3061 gattaactct agaggcagtg gagccactaa cttctccctg ttgaaacaag caggggatgt 3121 cgaagagaat cccgggccaa tggtgagcaa gggcgaggag gataacatgg ccatcatcaa 3181 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat 3241 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac 3301 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc 3361 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga 3421 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca 3481 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt 3541 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg 3601 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga 3661 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct 3721 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac 3781 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct 3841 gtacaagtaa cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca 3901 tgcatcgatg atctagagct ccgtctcaag ctgagctcgc tgatcagcct cgactgtgcc 3961 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 4021 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 4081 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 4141 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaacctg 4201 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 4261 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 4321 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 4381 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 4441 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 4501 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga 4561 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata 4621 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 4681 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca 4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 4801 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 4861 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 4921 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 4981 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 5041 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 5101 tccgaccctg ccgcttaccg catacctgtc cgcctttctc ccttcgggaa gcgtggcgct 5161 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 5221 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 5281 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 5341 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 5401 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 5461 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 5521 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 5581 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 5641 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 5701 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 5761 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 5821 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 5881 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 5941 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6001 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6061 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6121 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6181 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 6241 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 6301 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 6361 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 6421 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 6481 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 6541 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 6601 ttttcaatga ccgctttgga aaaacaaaga ctgtatttcc tggaaattaa tgtttattca 6661 ataaactgtg tattcagcta tatcacatag tggtgaggct gaaatgaggc gggaagaggc 6721 ggttggggct taattatatc aatttgggtg gccccacagc gcctccaagg cgccagtcct 6781 gttttgacaa gttgcctctg gaagcctcta caatgcctct cttctttttc tccagagtaa 6841 gcggaggcca ggggcccccg gcctctgctt aatactaaaa aaaacagctg ttgtcatagt 6901 aatgattggg tggaaacatt ccaggcctgg gtggagaggc tttttgcttc ctcttgcaaa 6961 accacactga cattccaggc ctgggtggag aggctttttg cttcctcttg caaaaccaca 7021 ctgccctctg gagggcagtt gcctagcaac taactaaaag aggatgtcgc acggccagct 7081 gcggtcagtt agtcacttcc tgcttaactg acttgacatt ttctatttta agagtcggga 7141 ggaaaattac tgtgttggag gccctccgcc atcttctgaa gctgaatcga attaacttgt 7201 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 7261 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 7321 tctggatctg atatcatcgt c //