LOCUS Exported 7576 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of nucleus localized PAmCherry1 and EGFP fusion protein. ACCESSION . VERSION . KEYWORDS pCaggs-NLS-PAmCherry1-GSS-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7576) AUTHORS Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F TITLE Versatile phenotype-activated cell sorting. JOURNAL Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 10.1126/sciadv.abb7438. Print 2020 Oct. PUBMED 33097540 REFERENCE 2 (bases 1 to 7576) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7576 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(175..1035) /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 798..817 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1036..1140) /gene="bla" /label=AmpR promoter enhancer 1172..1551 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 1827..2843 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 2767..2789 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2851..2870 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 3013..3033 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3034..3741 /codon_start=1 /product="photoactivatable monomeric derivative of DsRed fluorescent protein (Subach et al., 2009)" /label=PAmCherry1 /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(3179..3197) /label=mCherry-R /note="mCherry, reverse primer" CDS 3769..4488 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3769..4485 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3814..3835) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4075..4094) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4422..4443 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4603..4620) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4609..4833 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4879..5307 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4966..4985) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5176..5197 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5316..5336) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5321..5650 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5501..5636 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5563..5582 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5661..6260 /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(5661..5680) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6157..6177 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 6420..6541 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6457..6476) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6511..6530 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6590..6606) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6590..6606) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6603..6625) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6614..6630 /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(6638..6668) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6683..6704 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6821..6838) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6992..4) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7072..7091) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 61 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 1021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 1081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 1141 cacatttccc cgaaaagtgc cacctgacgt cgacattgat tattgactag ttattaatag 1201 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 1261 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 1321 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 1381 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 1441 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 1501 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg 1561 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat 1621 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggsgcgccrg 1681 gsgggggggg gggggggggg ggggggggsg gggsgaggcg gagaggtgcg gcggcagcca 1741 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct 1801 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc 1861 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg 1921 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc 1981 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg 2041 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct 2101 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca 2161 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg 2221 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt 2281 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt 2341 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc 2401 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggrgggc tcgggggagg 2461 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc 2521 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc 2581 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg 2641 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc 2701 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca 2761 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc 2821 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc 2881 attttggcaa agaattcctc gaggaattca ctcctcaggt gcaggctgcc tatcagaagg 2941 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctg catgagctag 3001 ccgccaacca tgcccaagaa gaagaggaaa gtcatggtga gcaagggcga ggaggataac 3061 atggccatca ttaaggagtt catgcgcttc aaggtgcaca tggaggggtc cgtgaacggc 3121 cacgtgttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 3181 aagctgaagg tgaccaaggg cggccccctg cccttcacct gggacatcct gagccctcag 3241 ttcatgtacg gctccaatgc ctacgtgaag caccccgccg acatccccga ctactttaag 3301 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga aattcgagga cggcggcgtg 3361 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg 3421 cgcggcacca acttcccctc cgacggcccc gtgatgcaga agaagaccat gggctgggag 3481 gccctctccg agcggatgta ccccgaggac ggcgccctga agggcgaggt caagcccaga 3541 gtgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag 3601 aagcccgtgc agctgcccgg cgcctacaac gtcaaccgca agctggacat caccagccac 3661 aacgaggact acaccatcgt ggagcagtac gagagagccg agggccgcca ctccaccggc 3721 ggcatggacg agctgtacaa gggttcttca ggttcatctg gatcttccat ggtgagcaag 3781 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac 3841 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc 3901 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc 3961 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc 4021 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac 4081 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc 4141 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac 4201 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg 4261 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag 4321 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc 4381 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc 4441 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aaagcttggt 4501 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca gcacagtggc 4561 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta 4621 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca 4681 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc 4741 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata 4801 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg 4861 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 4921 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 4981 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 5041 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 5101 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 5161 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 5221 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 5281 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg 5341 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 5401 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 5461 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 5521 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga 5581 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg 5641 cttttgcaaa aagctcccgc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg 5701 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 5761 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 5821 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 5881 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 5941 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 6001 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc 6061 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 6121 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 6181 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 6241 cccgcaagcc cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 6301 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 6361 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 6421 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 6481 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6541 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 6601 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 6661 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 6721 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 6781 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 6841 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 6901 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 6961 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 7021 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 7081 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 7141 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 7201 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 7261 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 7321 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 7381 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 7441 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 7501 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 7561 gcagaaaaaa aggatc //