LOCUS Exported 3769 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Transient expression of the ""TOP1"" construct, targeting the GLuc reporter, in mammalian cells. CMV/3' Box expression backbone.. ACCESSION . VERSION . KEYWORDS pCMV/3' Box_(GLuc)_TOP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3769) AUTHORS Shechner DM, Hacisuleyman E, Younger ST, Rinn JL TITLE Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display. JOURNAL Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 2015 Jun 1. PUBMED 26030444 REFERENCE 2 (bases 1 to 3769) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3769 /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 1034..1052 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(1084..1106) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 1200..1219 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1413..1435 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1427..1444 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1428..1444 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 1463..1841 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1842..2045 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1996..2016 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_RNA 2101..2176 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(2609..2625) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2609..2625) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2622..2644) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2633..2649 /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(2657..2687) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2702..2723 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2840..2857) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3011..3599) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3091..3110) /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 gccctttgcg cgtttcggtg atgacggtga 1081 aaacctctga cacatgcagc tcccggtcac agcttgtctg taagcggatg ccgggagcag 1141 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 1201 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 1261 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 1321 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 1381 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 1441 cagtgaattc gagctcggta ctacattgat tattgactag ttattaatag taatcaatta 1501 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 1561 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 1621 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 1681 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 1741 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 1801 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 1861 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 1921 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 1981 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 2041 gagctctctg gctaactaga gaacccactg cttactggcg atctagatac gactcactat 2101 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt 2161 ggcaccgagt cggtgctctt caacatccct cctctaaacc cggaattgcg ggaaaggggt 2221 caacagccgt tcagtaccaa gtctcagggg aaactttgag atggccttgc aaagggtatg 2281 gtaataagct gacggacatg gtcctaacac gcagccaagt cctaagtcaa cagtttttgg 2341 agcagacgat atggcgtcgc tcctttttcc agcagagcat atgggctccg tggtttttcc 2401 agcagacgat atggcgtcgc tggtttttgg agcagagcat atgggctcgc tcctttttgc 2461 agcagacgat atggcgtcgc tgctttttct gttgatatgg atgcagttca actttctgga 2521 gtttcaaaag tagactgtac gctaagggtc atatcttttt ttgtttggtt tgtgtcttgg 2581 ttggcgtctt aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 2641 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 2701 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 2761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 2821 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 2881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 2941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 3001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 3061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 3121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 3181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 3241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 3301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 3361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 3421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 3481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 3541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 3601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 3661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 3721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacag //