LOCUS Exported 4911 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Vector encoding the EL222 gene under constitutive promoter J23106.. ACCESSION . VERSION . KEYWORDS pEBLindv2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4911) AUTHORS Jayaraman P, Yeoh JW, Jayaraman S, Teh AY, Zhang J, Poh CL TITLE Cell-Free Optogenetic Gene Expression System. JOURNAL ACS Synth Biol. 2018 Apr 20;7(4):986-994. doi: 10.1021/acssynbio.7b00422. Epub 2018 Apr 4. PUBMED 29596741 REFERENCE 2 (bases 1 to 4911) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4911 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 70..89 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(286..1164) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1191..1475 /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 1398..1417 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 1546..1557 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" terminator 2274..2345 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2361..2388 /label=T7Te terminator /note="phage T7 early transcription terminator" RBS 2459..2470 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 2477..3154 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 3187..3258 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3274..3301 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(3459..4047) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3539..3558) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 4135..4229 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(4260..143) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4371..4390) /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt 61 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg 121 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat 181 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc 241 ttcccaacct taccagaggg cgccccagct ggcaattccg acgtcttatg acaacttgac 301 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt 361 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac 421 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc 481 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga 541 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg 601 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc 661 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc 721 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg 781 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt 841 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg 901 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc 961 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag 1021 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt 1081 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag 1141 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa 1201 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac 1261 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac 1321 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca 1381 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg attctacctg 1441 acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg aattcaaaag 1501 atcttttacg gctagctcag tcctaggtat agtgctagcc tagagaaaga ggagaaatac 1561 tagatgttgg atatgggaca agatcggccg atcgatggaa gtggggcacc cggggcagac 1621 gacacacgcg ttgaggtgca accgccggcg cagtgggtcc tcgacctgat cgaggccagc 1681 ccgatcgcat cggtcgtgtc cgatccgcgt ctcgccgaca atccgctgat cgccatcaac 1741 caggccttca ccgacctgac cggctattcc gaagaagaat gcgtcggccg caattgccga 1801 ttcctggcag gttccggcac cgagccgtgg ctgaccgaca agatccgcca aggcgtgcgc 1861 gagcacaagc cggtgctggt cgagatcctg aactacaaga aggacggcac gccgttccgc 1921 aatgccgtgc tcgttgcacc gatctacgat gacgacgacg agcttctcta tttcctcggc 1981 agccaggtcg aagtcgacga cgaccagccc aacatgggca tggcgcgccg cgaacgcgcc 2041 gcggaaatgc tcaagacgct gtcgccgcgc cagctcgagg ttacgacgct ggtggcatcg 2101 ggcttgcgca acaaggaagt ggcggcccgg ctcggcctgt cggagaaaac cgtcaagatg 2161 caccgcgggc tggtgatgga aaagctcaac ctgaagacca gcgccgatct ggtgcgcatt 2221 gccgtcgaag ccggaatcta aggatccaaa ctcgagtaag gatctccagg catcaaataa 2281 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg 2341 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tataggtagc 2401 ctttagtcca tgttagcgaa gaaaatggtt tgttatagtc gaataaatat gtctagagaa 2461 agaggagaaa tactagatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa 2521 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg 2581 tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc 2641 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca 2701 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg 2761 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga 2821 cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt 2881 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg 2941 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc 3001 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac 3061 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga 3121 acgtgctgaa ggtcgtcact ccaccggtgc ttaaggatcc aaactcgagt aaggatctcc 3181 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt 3241 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc 3301 gtttatacct agggcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 3361 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 3421 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 3481 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 3541 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 3601 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 3661 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 3721 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 3781 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 3841 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 3901 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 3961 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 4021 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 4081 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactagtgc ttggattctc 4141 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga 4201 ggtcattact ggatctatca acaggagtcc aagcgagctc tcgaacccca gagtcccgct 4261 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata 4321 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg 4381 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat 4441 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg 4501 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg 4561 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta 4621 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc 4681 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga 4741 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca 4801 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc 4861 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac a //