LOCUS Exported 8309 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector that contains an optimized S. pyogenes sgRNA targeting human telomeres. ACCESSION . VERSION . KEYWORDS pSLQ1651-sgTelomere(F+E) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8309) AUTHORS Chen B, Gilbert LA, Cimini BA, Schnitzbauer J, Zhang W, Li GW, Park J, Blackburn EH, Weissman JS, Qi LS, Huang B TITLE Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR/Cas System. JOURNAL Cell. 2013 Dec 19;155(7):1479-91. doi: 10.1016/j.cell.2013.12.001. PUBMED 24360272 REFERENCE 2 (bases 1 to 8309) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8309 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(40..59) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 231..610 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 612..810 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 828..1008 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1055..1180 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1677..1910 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2095..2139 /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 2437..2554 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2807..2840 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 2844..2864 /label=mU6-F /note="Mouse U6 promoter, forward primer" enhancer 3159..3462 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3463..3666 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 3700..4296 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3700..3719) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4196..4216 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 4306..4359 /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" regulatory 4366..4375 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4372..5082 /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(4517..4535) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 4785..4804 /label=mCherry-F /note="mCherry, forward primer" protein_bind 5102..5135 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5191..5779 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5244..5264) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5662..5673) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5782..5798) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5783..5799) /label=pBluescriptKS /note="For pBluescript vector" LTR 6308..6488 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(6550..7138) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6630..6649) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7309..8169) /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 7932..7951 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8170..8274) /gene="bla" /label=AmpR promoter ORIGIN 1 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc 61 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 121 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 181 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt 241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 781 gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt 841 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 901 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 961 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa 1021 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc 1081 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac 1141 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat 1201 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa 1261 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga 1321 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc 1381 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat 1441 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa 1501 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg 1561 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1621 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1681 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1741 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1801 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1861 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1921 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1981 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 2041 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 2101 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 2161 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 2221 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 2281 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa 2341 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg 2401 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg 2461 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga 2521 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga 2581 tccagtttgg ttagtaccgg gcccgctcta gagatccgac gcgccatctc taggcccgcg 2641 ccggccccct cgcacggact tgtgggagaa gctcggctac tcccctgccc cggttaattt 2701 gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac agataatctg 2761 ttctttttaa tactagctac attttacatg ataggcttgg atttctataa cttcgtatag 2821 catacattat acgaagttat aaacagcaca aaaggaaact caccctaact gtaaagtaat 2881 tgtgtgtttt gagactataa gtatcccttg gagaaccacc ttgttggtta gggttagggt 2941 tagggttagt ttaagagcta tgctggaaac agcatagcaa gtttaaataa ggctagtccg 3001 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tctcgagtac taggatccat 3061 taggcggccg cgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt 3121 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 3181 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 3241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 3301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 3361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 3421 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag 3481 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 3541 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 3601 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 3661 agagctggtt tagtgaaccg tcagatccgc tagcccacca tgaccgagta caagcccacg 3721 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc 3781 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc 3841 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 3901 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 3961 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4021 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 4081 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 4141 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 4201 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 4261 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggat cgggagaggg cagaggaagt 4321 ctgctaacat gcggtgacgt cgaggagaat cctggcccac cggtcgccac catggtgagc 4381 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg 4441 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac 4501 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg 4561 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac 4621 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac 4681 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc 4741 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag 4801 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag 4861 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag 4921 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag 4981 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag 5041 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagaattcgt cgagggacct 5101 aataacttcg tatagcatac attatacgaa gttatacatg tttaagggtt ccggttccac 5161 taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 5221 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 5281 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 5341 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 5401 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 5461 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 5521 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 5581 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 5641 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 5701 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 5761 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg 5821 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca 5881 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta 5941 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 6001 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt 6061 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg 6121 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa 6181 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt 6241 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga 6301 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg 6361 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg 6421 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat 6481 ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 6781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 6841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 6901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 6961 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 7021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 7801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 7861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 7921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 7981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8281 tccccgaaaa gtgccacctg acgtcgacg //