LOCUS Exported 6225 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBAD-RFP(EC) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6225) AUTHORS Pfleger BF, Fawzi NJ, Keasling JD TITLE Optimization of DsRed production in Escherichia coli: effect of ribosome binding site sequestration on translation efficiency. JOURNAL Biotechnol Bioeng. 2005 Dec 5. 92(5):553-8. PUBMED 16247774 REFERENCE 2 (bases 1 to 6225) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6225 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 128..844 /codon_start=1 /product="GFP variant optimized for excitation by UV light" /label=GFPuv /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(158..186) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 781..801 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind complement(1076..1093) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 1246..1332 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1424..1451 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 1470..1561 /gene="bla" /label=AmpR promoter CDS 1562..2422 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1780..1799) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2464..2919 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2551..2570) /label=F1ori-R /note="F1 origin, reverse primer" rep_origin 3030..3618 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3519..3538 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3772..3789 /label=L4440 /note="L4440 vector, forward primer" primer_bind 3955..3977 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4116..4135 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(4315..5193) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 5220..5504 /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 5427..5446 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" primer_bind complement(5679..5697) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(5729..5749) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 5947..5966 /label=mCherry-F /note="mCherry, forward primer" primer_bind 6141..6164 /label=DsRed1-C /note="DsRed1, forward primer" ORIGIN 1 gatctgatcg ttgcgggcgg ggcgagagtc tcgccccgcc cgcgaccgcg gtgaaaatac 61 gagaatatta tttgtattga tctcctaggc ggggtaccgt attttggatg ataacgaggc 121 gcaaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt 181 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac 241 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc 301 aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat 361 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat 421 atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac 481 ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg 541 acacaaactg gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa 601 gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact 661 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa 721 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat 781 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa 841 ataagatgaa ttccagctga gcgccggtcg ctaccattac cagttggtct ggtgtcaaaa 901 ataataataa ccgggcaggg gggatctgtt aagttctcgt ctggtagaaa aacgctgctg 961 cgggtttttt tttgccttta gtaaattgaa ctgactttcg tcagttattc cttacccagc 1021 aatgcctgca ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga 1081 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta 1141 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg 1201 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag 1261 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg 1321 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg 1381 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg 1441 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat 1501 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 1561 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc 1621 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc 1681 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc 1741 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc 1801 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt 1861 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt 1921 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat 1981 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct 2041 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat 2101 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc 2161 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg 2221 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc 2281 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta 2341 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc 2401 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 2461 tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 2521 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 2581 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 2641 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg 2701 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 2761 gtggactctt gttccaaact tgaacaacac tcaaccctat ctcgggctat tcttttgatt 2821 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 2881 ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc 2941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 3001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 3061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 3121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 3181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 3241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 3301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 3361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 3421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 3481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 3541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 3601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 3661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 3721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 3781 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt 3841 atttcacacc gcatagggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc 3901 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga 3961 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag caaggagatg 4021 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc 4081 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca 4141 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc 4201 tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat 4261 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga 4321 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct 4381 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt 4441 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat 4501 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga 4561 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc 4621 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg 4681 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc 4741 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc 4801 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga 4861 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata 4921 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa 4981 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg 5041 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag 5101 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc 5161 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga 5221 agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt 5281 ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa 5341 agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga 5401 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga 5461 tcctacctga cgctttttat cgcaactctc tactgtttct ccatacgtcg acagctagcg 5521 tattttggat gataacgagg cgcaaaaaat ggcgagcagt gaggacatca tcaaggagtt 5581 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg 5641 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg 5701 cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt 5761 gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt 5821 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc 5881 ctccctgcag gacggctcct tcatctacaa ggtgaagttc atcggcgtga acttcccctc 5941 cgacggcccc gtaatgcaga agaagactat gggctgggag gcctccaccg agcgcctgta 6001 cccccgcgac ggcgtgctga agggcgagat ccacaaggcc ctgaagctga aggacggcgg 6061 ccactacctg gtggagttca agtccatcta catggccaag aagcccgtgc agctgcccgg 6121 ctactactac gtggactcca agctggacat cacctcccac aacgaggact acaccatcgt 6181 ggagcagtac gagcgcgccg agggccgcca ccacctgttc ctgta //