LOCUS pJL1-sf-GFP(PC,T7_tag) 3262 bp DNA circular UNA 14-JUL-2025 DEFINITION From PLD: sequencing completely. ACCESSION urn.local...6a-jeo046a VERSION urn.local...6a-jeo046a KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers source 1..3262 /mol_type="other DNA" /organism="synthetic DNA construct" /label="source" misc_feature 139..158 /standard_name="gRNA3-ori-" /label="gRNA3-ori-" promoter 630..648 /note="promoter for bacteriophage T7 RNA polymerase" /standard_name="T7 promoter" /modified_by="La Fee Bleue" /label="T7 promoter" RBS 696..701 /label="ribosome binding site" tag 722..739 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /modified_by="La Fee Bleue" /label="6xHis" protein_bind 749..766 /codon_start=1 /product="thrombin recognition and cleavage site" /transl_table=1 /translation="LVPRGS" /modified_by="La Fee Bleue" /label="thrombin site" tag 770..802 /codon_start=1 /note="promotes efficient translation in E. coli" /product="leader peptide from bacteriophage T7 gene 10" /transl_table=1 /translation="MASMTGGQQMG" /modified_by="La Fee Bleue" /label="T7 tag (gene 10 leader)" protein_bind 806..850 /note="multiple cloning site" /standard_name="MCS" /label="MCS binding site" gene 854..1567 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Péde lacq et al., 2006)" /transl_table=1 /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKD DGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNG IKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHM VLLEFVTAAGITHGMDELYK" /modified_by="La Fee Bleue" /label="superfolder GFP" terminator 1636..1683 /note="transcription terminator for bacteriophage T7 RNA polymerase" /standard_name="T7 terminator" /modified_by="La Fee Bleue" /label="T7 terminator" terminator 1845..1893 /note="central terminator from bacteriophage fd (Otsuk a and Kunisawa, 1982)" /label="fd terminator" CDS complement(2057..2872) /codon_start=1 /gene="aph(3')-Ia" /note="confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes" /product="aminoglycoside phosphotransferase" /transl_table=1 /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLY GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*" /label="KanR" rep_origin 2994..320 /snapgene_direction="RIGHT" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS 3030..320 /standard_name="RNA II" /label="RNA II" CDS complement(3033..3140) /standard_name="RNAI" /label="RNAI" misc_feature 3051..3070 /standard_name="gRNA1-ori+" /label="gRNA1-ori+" misc_feature 3140..3184 /standard_name="putative RNAI promoter" /label="putative RNAI promoter" misc_feature 3226..3245 /standard_name="gRNA2-ori-" /label="gRNA2-ori-" ORIGIN 1 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 61 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 301 gggggcggag cctatggaaa aacgccagca acgcgatccc gcgaaatgct catgagcgga 361 gaacgagatg acgttggagg ggcaaggtcg cgctgattgc tggggcaaca cgtggagcgg 421 atcggggatt gtctttcttc agctcgctga tgatatgctg acgctcaatg ccgtttggcc 481 tccgactaac gaaaatcccg catttggacg gctgatccga ttggcacggc ggacggcgaa 541 tggcggagca gacgctcgtc cgggggcaat gagatatgaa aaagcctgaa ctcaccgcga 601 cgtatcgggc cctggccagg atctgatatt aatacgactc actataggga gaccacaacg 661 gtttccctat agaaataatt ttgtttaact ttaagaagga gaatatacca tgggcagcag 721 ccatcatcat catcatcaca gcagcggcct ggtgccgcgc ggcagccata tggctagcat 781 gactggtgga cagcaaatgg gtcgcggatc cgaattcgag ctccgtcgac aagcttgcgg 841 ccgcactcga ggtatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt 901 ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga 961 cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggtacc 1021 ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga 1081 ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg 1141 cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg 1201 cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat 1261 cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa 1321 acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt 1381 gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc 1441 agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga 1501 tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact 1561 gtacaaatga gtcgaccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc 1621 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt 1681 ttgctgaaag ccaattctga gatcccgcaa aagcggcctt tgactccctg caagcctcag 1741 cgaccgaata tatcggttat gcgtgggcga tggttgttgt cattgtcggc gcaactatcg 1801 gtatcaagct gtttaagaaa ttcacctcga aagcaagctg ataaaccgat acaattaaag 1861 gctccttttg gagccttttt ttttggagat tttcaacgtg aaaaaattat tattcgcaat 1921 tcctttagtt gttcctttct attctcactc cgctgaaact gttgaaagtt gtttagcaaa 1981 acctcataca gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcctct 2041 agctagagtc gaccggttag aaaaactcat cgagcatcaa atgaaactgc aatttattca 2101 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact 2161 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc 2221 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat 2281 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga 2341 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt 2401 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat 2461 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt 2521 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg 2581 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa 2641 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt 2701 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg 2761 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt 2821 tggaatttaa tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc 2881 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt 2941 gtgcaatgta acatcagaga ttttgagaca caacgtgaga tcaaaggatc ttcttgagat 3001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 3061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 3121 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 3181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 3241 ggcgataagt cgtgtcttac cg //