LOCUS Exported 5374 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses GFP with 3 UAG codons at the amino acid position 3, 151, and 153. ACCESSION . VERSION . KEYWORDS pGLO-GFP-3UAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5374) AUTHORS Schwessinger B, Li X, Ellinghaus TL, Chan LJ, Wei T, Joe A, Thomas N, Pruitt R, Adams PD, Chern MS, Petzold CJ, Liu CC, Ronald PC TITLE A second-generation expression system for tyrosine-sulfated proteins and its application in crop protection. JOURNAL Integr Biol (Camb). 2016 Apr 18;8(4):542-5. doi: 10.1039/c5ib00232j. Epub 2015 Nov 27. PUBMED 26611838 REFERENCE 2 (bases 1 to 5374) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5374 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(96..974) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1001..1285 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 1208..1227 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 1312..1334 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1351..2064 /codon_start=1 /product="Cycle 3 GFP (Crameri et al., 1996)" /label=Cycle 3 GFP /note="mammalian codon-optimized" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSH*V*ITADKQKNGIKAN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" primer_bind complement(1378..1406) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2001..2021 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" misc_feature 2066..2122 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(2155..2172) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2324..2410 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2502..2529 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 2547..2638 /gene="bla" /label=AmpR promoter CDS 2639..3499 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2857..2876) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3541..3996 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3628..3647) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3838..3859 /label=F1ori-F /note="F1 origin, forward primer" rep_origin 4107..4695 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4596..4615 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4849..4866 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4881..5021 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4947..4966) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5107..5129 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5268..5287 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac 61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca 121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta 181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata 241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag 301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag 361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg 421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct 481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc 541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc 601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca 661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga 721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa 781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata 841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc 901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt 961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat 1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta 1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt 1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca 1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta 1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagaaataa ttttgtttaa 1321 ctttaagaag gagatataca tatggcttag agcaaaggag aagaactttt cactggagtt 1381 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga 1441 gagggtgaag gtgatgctac atacggaaag cttaccctta aatttatttg cactactgga 1501 aaactacctg ttccatggcc aacacttgtc actactttct cttatggtgt tcaatgcttt 1561 tcccgttatc cggatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt 1621 tatgtacagg aacgcactat atctttcaaa gatgacggga actacaagac gcgtgctgaa 1681 gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaaaggtat tgattttaaa 1741 gaagatggaa acattctcgg acacaaactc gagtacaact ataactcaca ctaggtatag 1801 atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacatt 1861 gaagatggat ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc 1921 cctgtccttt taccagacaa ccattacctg tcgacacaat ctgccctttc gaaagatccc 1981 aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat 2041 ggcatggatg agctctacaa ataatgaatt cgagctcggt acccggggat cctctagagt 2101 cgacctgcag gcatgcaagc ttggctgttt tggcggatga gagaagattt tcagcctgat 2161 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag 2221 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg 2281 tagtgtgggg tcccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc 2341 tcagtgcaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag 2401 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg 2461 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga 2521 tggccttttt gcgtttctac aaactctttg tttatttttc taaatacatt caaatatgta 2581 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 2641 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 2701 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 2761 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 2821 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 2881 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 2941 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 3001 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 3061 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 3121 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 3181 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 3241 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 3301 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 3361 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 3421 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 3481 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 3541 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 3601 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 3661 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 3721 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 3781 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 3841 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat 3901 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 3961 acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt 4021 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 4081 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 4141 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 4201 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta 4261 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 4321 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 4381 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 4441 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat 4501 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 4561 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 4621 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 4681 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 4741 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 4801 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 4861 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 4921 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 4981 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac 5041 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 5101 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc 5161 agcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg 5221 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg 5281 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg 5341 tagaggatct aattctcatg tttgacagct tatc //