LOCUS Exported 6816 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of mGold in mammalian cells. ACCESSION . VERSION . KEYWORDS pCaggs-mGold SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6816) 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 6816) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6816 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 15..37 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 99..118 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 251..260 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 257..976 /codon_start=1 /product="photostable derivative of mVenus (Lee et al., 2020)" /label=mGold /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(302..323) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(563..582) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 910..931 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1091..1108) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1097..1321 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1367..1795 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1454..1473) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1664..1685 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1804..1824) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1809..2138 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1989..2124 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2051..2070 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2149..2748 /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(2149..2168) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2645..2665 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 2908..3029 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2945..2964) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2999..3018 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3078..3094) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3078..3094) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3091..3113) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3102..3118 /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(3126..3156) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3171..3192 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3309..3326) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3480..4068) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3560..3579) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4239..5099) /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 4862..4881 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5100..5204) /gene="bla" /label=AmpR promoter enhancer 5236..5615 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 5891..91 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" ORIGIN 1 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa 61 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct 121 gtctcatcat tttggcaaag aattcctcga ggaattcact cctcaggtgc aggctgccta 181 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctgca 241 tgagctagcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 421 cgtgccctgg cccaccctcg tgaccagcct gggctacggc ctgcagtgct tcgcccgcta 481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 661 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc 721 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg 781 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 841 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa 901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 961 cgagctgtac aagtaatgaa agcttggtac cgagctcgga tccactagtc cagtgtggtg 1021 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc 1081 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 1141 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 1201 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 1261 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 1321 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 1381 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 1441 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 1501 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 1561 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 1621 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 1681 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 1741 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 1801 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 1861 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 1921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 1981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 2041 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 2101 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccgcat gaccgagtac 2161 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 2221 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 2281 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 2341 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 2401 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 2461 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 2521 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 2581 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 2641 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 2701 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc 2761 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 2821 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 2881 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 2941 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 3001 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 3061 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 3121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 3181 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 3241 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 3301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 3361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 4201 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 4261 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 4321 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 4381 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 4441 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 4501 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 4561 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 4621 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 4681 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 4741 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 4801 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 4861 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 4921 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 4981 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 5041 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 5101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 5161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 5221 gtgccacctg acgtcgacat tgattattga ctagttatta atagtaatca attacggggt 5281 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc 5341 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag 5401 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc 5461 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg 5521 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc 5581 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct 5641 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat 5701 tattttgtgc agcgatgggg gcgggggggg ggggggggcg ccrggsgggg gggggggggg 5761 gggggggsgg ggsggggsgr ggcggagagg tgcggcggca gccaatcaga gcggcgcgct 5821 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc 5881 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc 5941 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg 6001 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg 6061 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg 6121 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg 6181 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag 6241 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt 6301 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc 6361 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg 6421 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc 6481 gggcggggcg gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg 6541 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg 6601 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg 6661 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat 6721 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg 6781 gggctgtccg cggggggacg gctgccttcg gggggg //