LOCUS Exported 3460 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Provides the 4xLexA Operator fused to CaMV 35s minimal promoter as a level 0 GoldenBraid part. ACCESSION . VERSION . KEYWORDS pOpLexA::mini35s (GB0733) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3460) AUTHORS Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D TITLE GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. JOURNAL Plant Physiol. 2013 Jul;162(3):1618-31. PUBMED 23669743 REFERENCE 2 (bases 1 to 3460) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3460 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(82..128) /label=minimal CaMV 35S promoter /note="minimal 35S promoter from cauliflower mosaic virus" promoter complement(510..528) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(511..528) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(546..562) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(546..562) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(559..581) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 570..586 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(594..624) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 639..660 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(777..794) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(948..1536) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1028..1047) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1707..2567) /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 2330..2349 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2568..2672) /gene="bla" /label=AmpR promoter rep_origin complement(2750..3205) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2887..2908) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3099..3118 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3331..3353 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3345..3362 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3346..3362 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3369..3388 /label=T7 /note="T7 promoter, forward primer" promoter 3369..3387 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 3418..3427 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 3422..3431 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 ctcgcatttc gactagaata gtaaattgta atgttgtttg ttgtttgttt tgttgtggta 61 attgttgtaa aaataccgga gtcctctcca aatgaaatga acttccttat atagaggaag 121 ggtcttgcga aggggagcat attacatggc attaccacca tatacatatc catatacata 181 tccatatcta atcttacctc gactgctgta tataaaacca gtggttatat gtacagtact 241 gctgtatata aaaccagtgg ttatatgtac agtacgtcga ctgctgtata taaaaccagt 301 ggttatatgt acagtactgc tgtatataaa accagtggtt atatgtacag tacgtcgagg 361 ggatgataat gcgattagtt ttttagcctc gacctgcagc ccaagctgtc acgagaccac 421 tcatcgccca atcactagtg aattcgcggc cgcctgcagg tcgaccatat gggagagctc 481 ccaacgcgtt ggatgcatag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa 541 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 601 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 1021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 1081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 1141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 1201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 1261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 1321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 1381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 1441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 1501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 1561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 1621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 1681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 1741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 1801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgcga cccacgctca 1861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 1921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 1981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 2041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 2101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 2161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 2221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 2281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 2341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 2401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 2461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 2521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 2581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 2641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat 2701 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc 2761 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 2821 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 2881 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 2941 cgaaaaaccg tctatcaggg cgctggccca ctacgtgaac catcacccta atcaagtttt 3001 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 3061 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 3121 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 3181 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag 3241 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 3301 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 3361 gtgaattgta atacgactca ctatagggcg aattgggccc gacgtcgcat gctcccggcc 3421 gccatggcgg ccgcgggaat tcgattgggc gatgagtggt //