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 <i>E. coli</i>"
                     /product="leader peptide from bacteriophage T7 gene
                     <i>10</i>"
                     /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 (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/16369541""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/16369541"">Péde
                     lacq et al., 2006</a>)"
                     /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 (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/6215541""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/6215541"">Otsuk
                     a and Kunisawa, 1982</a>)"
                     /label="fd terminator"
     CDS             complement(2057..2872)
                     /codon_start=1
                     /gene="<i>aph(3')-Ia</i>"
                     /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
//
