LOCUS Exported 7994 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pRipTAL plasmid compatible with Golden Gate TALEN cloning system. For cloning of megaTALs or other TAL-fusions and expression in mammalian cells. Meganuclease/fusion is cloned between Xba-Sal sites.. ACCESSION . VERSION . KEYWORDS pCVL RipTAL(Nd154C63) SFFV Xba-Sal linker SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7994) TITLE TALEN Golden Gate vectors JOURNAL Unpublished REFERENCE 2 (bases 1 to 7994) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7994 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 9..235 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 236..416 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 463..588 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1080..1313 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1498..1542 /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 1809..1926 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2621..3022 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" CDS 3027..3728 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MASELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY CDLPSKLGHKLN" CDS 3732..3785 /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" CDS 3792..3818 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 3822..3842 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(3867..4550) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGG TGKLPSQPIQGLDCWPLSVLGHCGDLGGGQPQRGGSKCGSHRQQGLERQISAPLPGDIH YVSASQFWQGGGELGVVPSDHAQTLSPPRNSADISVSKTMSDQGLMMLGNRASHFRLNF FLLLL" primer_bind 4400..4422 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4414..4431 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4415..4431 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 4441..4460 /label=T7 /note="T7 promoter, forward primer" promoter 4441..4459 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4507..4527) /label=T3 /note="T3 promoter, forward primer" promoter complement(4507..4525) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4546..4562) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4546..4562) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4559..4581) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4570..4586 /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(4594..4624) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4639..4660 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." protein_bind 5533..5566 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(5771..5788) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5777..6001 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6074..6091) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6245..6833) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6325..6344) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7004..7864) /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 7627..7646 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7865..7969) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtcaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta 61 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg 121 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac 181 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc 241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg 421 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact 481 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa 541 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg 601 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat 661 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc 721 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag 781 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat 841 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac 901 aaaagtaaga ccaccgcaca gcaagcggcc ctgatcttca gacctggagg aggagatatg 961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1381 aatgaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat 1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta 1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa 1921 aattttatcg attacgcgtg tggcatctga agcaccacca gcgagcgaga gctagagaga 1981 aggaaagcca ccgacttcac cgcctccgag ctgctccggg tcgcgggtct gcagcgtcta 2041 cggccctccg cgcctacagc tcaagccaca tccgaagggg gagggagccg ggagctgcgc 2101 gcggggccgc cggggggagg ggtggcaccg cccacgccgg gcggccacga agggcggggc 2161 agcgggcgcg cgcccggcgg ggggaggggc cgcgcgccgc gcccgctggg aattggggcc 2221 ctagggggag ggcggaggcg ccgacgaccg cggcacttac cgttcgcggc gtggcgcccg 2281 gtggtcccca aggggaggga agggggaggc ggggcgagga cagtgaccgg agtctcctca 2341 gcggtggctt ttctgcttgg cagcctcagc ggctggcgcc aaaaccggac tccgcccact 2401 tcctcgcccc tgcggtgcga gggtgtggaa tcctccagac gctgggggag ggggagttgg 2461 gagcttaaaa actagtaccc ctttgggacc actttcagca gcgaactctc ctgtacacca 2521 ggggtcagtt ccacagacgc gggccagggg tgggtcattg cggcgtgaac aataatttga 2581 ctagaagttg attcgggtgt ttccggaatt cctagctgca gtaacgccat tttgcaaggc 2641 atggaaaaat accaaaccaa gaatagagaa gttcagatca agggcgggta catgaaaata 2701 gctaacgttg ggccaaacag gatatctgcg gtgagcagtt tcggccccgg cccggggcca 2761 agaacagatg gtcaccgcag tttcggcccc ggcccgaggc caagaacaga tggtccccag 2821 atatggccca accctcagca gtttcttaag acccatcaga tgtttccagg ctcccccaag 2881 gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt 2941 tcgcgcgctt ctgcttcccg agctctataa aagagctcac aacccctcac tcggcgcgcc 3001 agtcctccga cagactgagt cgcccgatgg ctagcgagct gattaaggag aacatgcaca 3061 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg 3121 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc 3181 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca 3241 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg 3301 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc 3361 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc 3421 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacactg taccccgctg 3481 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga 3541 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg 3601 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg 3661 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca 3721 agcttaattc agagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatccgg 3781 gcccctgcag gtatccatat gatgtcccag attatgcgcc acctaagaag aaacgcaaag 3841 tcgtagacct cagaaccctc ggctattcac agcagcagca ggaaaaaatt aagcctaaag 3901 tgcgaagcac ggttgcccag catcatgagg ccttggtcgg acatggtttt actcacgctg 3961 atatctgcag aatttctcgg cggagacaga gtttgcgcgt ggtcgctcgg aactaccccg 4021 agctcgccgc cgccctgcca gaactgactc gcgctcacat agtggatatc gcccggcagc 4081 ggagcggaga tctggcgctc caagccctgc tgcctgtggc taccgcactt actgccgccc 4141 cgctgcggct gtccgcctcc cagatcgcca cagtggccca gtacggagag cggccagcaa 4201 tccaagccct gtataggctg cgaaggaagc ttaccggtgc ccccctggag acgggcgccg 4261 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 4321 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt 4381 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg 4441 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cctccagctt 4501 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc 4561 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4621 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4681 cgctttccac cggtcgtctc caacgaccac ctcgtcgcct tggcctgcct cggcggacgt 4741 cctgccatgg atgcagtgaa aaagggattg ccgcacgcgc cggaattgat cagaagagtc 4801 aatcgccgta ttggcgaacg cacgtcccat cgcgttgcga tatctagatg acgtagtgat 4861 ctgatctcag tcgaccgagc atcttaccgc catttatacc catatttgtt ctgtttttct 4921 tgatttgggt atacatttaa atgttaatag aacaaaatgg tggggcaatc atttacattt 4981 ttagggatat gtaattacta gttcaggtgt attgccacaa gacaaacatg ttaagaaact 5041 ttcccgttat ttacgctctg ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag 5101 attgactgat attcttaact atgttgctcc ttttacgctg tgtggatatg ctgctttata 5161 gcctctgtat ctagctattg cttcccgtac ggctttcgtt ttctcctcct tgtataaatc 5221 ctggttgctg tctcttttag aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg 5281 ctctgtgttt gctgacgcaa cccccactgg ctggggcatt gccaccacct gtcaactcct 5341 ttctgggact ttcgctttcc ccctcccgat cgccacggca gaactcatcg ccgcctgcct 5401 tgcccgctgc tggacagggg ctaggttgct gggcactgat aattccgtgg tgttgtcatc 5461 gaattcggta cctttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga 5521 agacaagata tcataacttc gtatagcata cattatacga agttataatt tatttgtgaa 5581 atttgtgatg ctattgcttt atttgtaacc atatgtttat ttgtgaaatt tgtgatgcta 5641 ttgctttatt tgtaaccatt gctttttgct tgtactgggt ctctctggtt agaccagatc 5701 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg 5761 cctcgaccag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 5821 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 5881 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 5941 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 6001 gcctgcagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6061 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 6181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 6241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 6301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 6361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 6421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 6481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 6541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 6601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 6661 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 6721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 6781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 6841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 6901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 6961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 7321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 7381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 7441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 7501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 7561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 7621 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 7681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 7741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 7801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 7861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 7921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 7981 gaaaagtgcc acct //