LOCUS Exported 4297 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of a fluorescent protein, tandem dimer Tomato (tdTomato). ACCESSION . VERSION . KEYWORDS pNCS tdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4297) AUTHORS Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY TITLE Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. JOURNAL Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21. PUBMED 15558047 REFERENCE 2 (bases 1 to 4297) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4297 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 20..39 /label=T7 /note="T7 promoter, forward primer" promoter 20..38 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 70..92 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 112..129 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 133..165 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 169..192 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" regulatory 199..208 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 205..1635 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" primer_bind complement(1698..1716) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1712..1759 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1856..2311 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1943..1962) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2153..2174 /label=F1ori-F /note="F1 origin, forward primer" promoter 2337..2441 /gene="bla" /label=AmpR promoter CDS 2442..3302 /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 complement(2660..2679) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3473..4061 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3962..3981 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4215..4232 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct 61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat 121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac 181 gatgacgata aggatcccgc caccatggtg agcaagggcg aggaggtcat caaagagttc 241 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc 301 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc 361 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg 421 tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc 481 aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc 541 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc 601 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac 661 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc 721 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc 781 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg 841 gaacagtacg agcgctccga gggccgccac cacctgttcc tggggcatgg caccggcagc 901 accggcagcg gcagctccgg caccgcctcc tccgaggaca acaacatggc cgtcatcaaa 961 gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc 1021 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc 1081 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc 1141 aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag 1201 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag 1261 gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc 1321 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc 1381 ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac 1441 ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg 1501 cccggctact actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc 1561 atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac 1621 gagctgtaca agtaagaatt cgaagcttga tccggctgct aacaaagccc gaaaggaagc 1681 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg 1741 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatctggcg taatagcgaa 1801 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg 1861 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 1921 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 1981 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 2041 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 2101 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 2161 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 2221 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 2281 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg 2341 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 2401 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 2461 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 2521 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 2581 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 2641 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 2701 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 2761 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 2821 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 2881 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 2941 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 3001 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 3061 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 3121 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 3181 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 3241 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 3301 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 3361 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 3421 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 3481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 3541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 3601 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 3661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 3721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 3781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 3841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 3901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 3961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 4021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 4081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 4141 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc 4201 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac 4261 cgcctctccc cgcgcgttgg ccgattcatt aatgcag //