LOCUS Exported 7208 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Destination vector for domain insertion. ACCESSION . VERSION . KEYWORDS pTALE-STAR-C_RFP entry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7208) 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 7208) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7208 /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" misc_feature 2671..2692 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" protein_bind 2793..2814 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2829..2859 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2867..2883 /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)." RBS 2901..2912 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 2919..3596 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 3641..3712 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3728..3755 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 3762..3782 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(3815..3832) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3821..4045 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4092..4108) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4092..4108) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4105..4127) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4116..4132 /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(4140..4170) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4185..4206 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4265..4460 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4311..4446 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4373..4392 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4466..4600 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4668..4685) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4839..5427) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4919..4938) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5598..6458) /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 6221..6240 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 7061..7195 /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 ctcacgacgg 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 ggacagtaga gacgccattg gaattcgcgg ccgcttctag agcaatacgc 2701 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 2761 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 2821 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 2881 caatttcaca catactagag aaagaggaga aatactagat ggcttcctcc gaagacgtta 2941 tcaaagagtt catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg 3001 aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag 3061 ttaccaaagg tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg 3121 gttccaaagc ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc 3181 cggaaggttt caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta 3241 cccaggactc ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca 3301 acttcccgtc cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg 3361 aacgtatgta cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga 3421 aagacggtgg tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc 3481 agctgccggg tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact 3541 acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaataac 3601 gctgatagtg ctagtgtaga tcgctactag agccaggcat caaataaaac gaaaggctca 3661 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag 3721 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagtag cggccgctgc 3781 agtcagaccg tctcaagctg agctcgctga tcagcctcga ctgtgccttc tagttgccag 3841 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 3901 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 3961 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 4021 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaacctgcag cccaagcttg 4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg 4261 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 4381 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 4441 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata 4501 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 4561 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc 4621 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 4681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 4861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 4921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 4981 cttaccgcat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca 5041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5221 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5461 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 5521 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 5581 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 5641 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 5701 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 5761 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 5821 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 5881 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 5941 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6001 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6061 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 6121 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 6181 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 6241 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 6301 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 6361 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 6421 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgaccg 6481 ctttggaaaa acaaagactg tatttcctgg aaattaatgt ttattcaata aactgtgtat 6541 tcagctatat cacatagtgg tgaggctgaa atgaggcggg aagaggcggt tggggcttaa 6601 ttatatcaat ttgggtggcc ccacagcgcc tccaaggcgc cagtcctgtt ttgacaagtt 6661 gcctctggaa gcctctacaa tgcctctctt ctttttctcc agagtaagcg gaggccaggg 6721 gcccccggcc tctgcttaat actaaaaaaa acagctgttg tcatagtaat gattgggtgg 6781 aaacattcca ggcctgggtg gagaggcttt ttgcttcctc ttgcaaaacc acactgacat 6841 tccaggcctg ggtggagagg ctttttgctt cctcttgcaa aaccacactg ccctctggag 6901 ggcagttgcc tagcaactaa ctaaaagagg atgtcgcacg gccagctgcg gtcagttagt 6961 cacttcctgc ttaactgact tgacattttc tattttaaga gtcgggagga aaattactgt 7021 gttggaggcc ctccgccatc ttctgaagct gaatcgaatt aacttgttta ttgcagctta 7081 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 7141 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgata 7201 tcatcgtc //