LOCUS Exported 5082 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Transient expression of non-aggregating delta-FG mutant, fused to mCherry in mammalian cells, under a hUBC promoter, HA tag. ACCESSION . VERSION . KEYWORDS pUBC_HA_PP7-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5082) AUTHORS Maass PG, Barutcu AR, Shechner DM, Weiner CL, Mele M, Rinn JL TITLE Spatiotemporal allele organization by allele-specific CRISPR live-cell imaging (SNP-CLING). JOURNAL Nat Struct Mol Biol. 2018 Jan 8. pii: 10.1038/s41594-017-0015-3. doi: 10.1038/s41594-017-0015-3. PUBMED 29343869 REFERENCE 2 (bases 1 to 5082) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5082 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1..861) /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 624..643 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(862..966) /gene="bla" /label=AmpR promoter primer_bind 1056..1075 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1269..1291 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1283..1300 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1284..1300 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2377..2396 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" regulatory 2558..2567 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2570..2590 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2597..2623 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 3035..3745 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGREFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(3180..3198) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3448..3467 /label=mCherry-F /note="mCherry, forward primer" primer_bind 3642..3665 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(3792..3811) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3846..3865 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3922..3938) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3922..3938) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3935..3957) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3946..3962 /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(3970..4000) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4015..4036 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4153..4170) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4324..4912) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4404..4423) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 1021 attaacctat aaaaataggc gtatcacgag ctatgcggca tcagagcaga ttgtactgag 1081 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 1141 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1201 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1261 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacacg 1321 tatgtcgagg taggcgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc 1381 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct 1441 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga 1501 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac 1561 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga 1621 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg 1681 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct 1741 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga 1801 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg 1861 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg 1921 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt 1981 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg 2041 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg 2101 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc 2161 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg 2221 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc 2281 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc 2341 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc 2401 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat 2461 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt 2521 ttttggcttt tttgttagac ctggaggtac cgccaccgcc accatggctc caaagaagaa 2581 gcgtaaggta ggcagctacc catacgatgt tccagattac gctggcagct ccatgtcaaa 2641 gacgatcgtg ctgtctgtag gtgaagccac acgcaccctc actgagatac agtccacggc 2701 cgataggcag atttttgagg agaaggtggg tccactggtg ggtaggcttc gactgacggc 2761 ctccctgagg caaaacggcg ctaaaacggc ttatcgcgtt aacctcaagc tggaccaggc 2821 cgatgtcgtt gatagtggac tgcccaaagt caggtacaca caggtgtgga gccacgatgt 2881 gaccatcgtg gctaactcaa cagaggcgtc ccggaagtct ctgtacgacc tgacaaagtc 2941 cctggttgcc acctctcagg tggaggatct ggtggttaat ctcgtgcccc tcggacgggg 3001 atccaagggc tcgggctcga cctcgggctc cggaatggtg agcaagggcg aggaggacaa 3061 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg 3121 ccgcgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc 3181 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctca 3241 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa 3301 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt 3361 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct 3421 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga 3481 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag 3541 gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct acaaggccaa 3601 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagctggaca tcacctccca 3661 caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc actccaccgg 3721 cggcatggac gagctgtaca agtaatgata agtcgacttg tttattgcag cttataatgg 3781 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 3841 tagttgtggt ttgtccaaac tcatcaatgt atcttattcg actgtttaaa cctgcaggca 3901 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 3961 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4021 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4081 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4141 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4201 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4261 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4321 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 4621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 4681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 4741 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 4801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 4861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 4921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 4981 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5041 taaatcaatc taaagtatat atgagtaaac ttggtctgac ag //