LOCUS Exported 3599 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Encodes the fluorescent protein amFP486/K68M for constitutive expression in E. coli.. ACCESSION . VERSION . KEYWORDS pThom77.3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3599) AUTHORS Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, Hughes TE, Drobizhev M TITLE Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than Enhanced Green Fluorescent Protein under Two-Photon Excitation. JOURNAL J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 10.1021/acs.jpclett.7b00960. Epub 2017 May 25. PUBMED 28530831 REFERENCE 2 (bases 1 to 3599) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3599 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 190..209 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 443..460 /label=L4440 /note="L4440 vector, forward primer" primer_bind 545..564 /label=T7 /note="T7 promoter, forward primer" promoter 545..563 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 647..669 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 689..706 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 710..742 /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 746..769 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" primer_bind complement(1525..1543) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1539..1586 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1683..2138 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1770..1789) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1980..2001 /label=F1ori-F /note="F1 origin, forward primer" promoter 2164..2268 /gene="bla" /label=AmpR promoter CDS 2269..3129 /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(2487..2506) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3300..289 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 61 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 361 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 421 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat 481 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct cgatcccgcg 541 aaattaatac gactcactat agggagacca caacggtttc cctctagaat caccgagttt 601 attcttgaca cctgatgcga tgaatgatat aataggaaag tactgttttg tttaacttta 661 agaaggagat atacatatgc ggggttctca tcatcatcat catcatggta tggctagcat 721 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atccggccct 781 gagcaacaag ttcatcggcg acgacatgaa gatgacctac cacatggacg gctgcgtgaa 841 cggccactac ttcaccgtga agggcgaggg caacggcaag ccctacgagg gcacccagac 901 cagcaccttc aaggtgacca tggccaacgg cggccccctg gccttcagct tcgacatcct 961 gagcaccgtg ttcatgtacg gcaaccggtg cttcaccgcc taccccacca gcatgcccga 1021 ctacttcaag caggccttcc ccgacggcat gagctacgag cggaccttca cctacgagga 1081 cggcggcgtg gccaccgcca gctgggagat cagcctgaag ggcaactgct tcgagcacaa 1141 gagcaccttc cacggcgtga acttccccgc cgacggcccc gtgatggcca agaagaccac 1201 cggctgggac cccagcttcg agaagatgac cgtgtgcgac ggcatcctga agggcgacgt 1261 gaccgccttc ctgatgctgc agggcggcgg caactaccgg tgccagttcc acaccagcta 1321 caagaccaag aagcccgtga ccatgccccc caaccacgtg gtggagcacc ggatcgcccg 1381 gaccgacctg gacaagggcg gcaacagcgt gcagctgacc gagcacgccg tggcccacat 1441 caccagcgtg gtgcccttct aagaattcga agcttgatcc ggctgctaac aaagcccgaa 1501 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct 1561 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tctggcgtaa 1621 tagcgaagag gccagcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 1681 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 1741 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 1801 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 1861 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 1921 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 1981 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 2041 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 2101 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat 2161 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 2221 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 2281 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 2341 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 2401 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 2461 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 2521 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 2581 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 2641 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 2701 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 2761 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 2821 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 2881 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 2941 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 3001 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 3061 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 3121 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 3181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 3241 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 3301 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 3361 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 3421 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 3481 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 3541 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataa //