LOCUS Exported 3144 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway entry clone encoding human GABARAP lacking stop codon, suitable for C-terminal tagging. ACCESSION . VERSION . KEYWORDS pDONR223 GABARAP no-stop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3144) AUTHORS Agrotis A, Pengo N, Burden JJ, Ketteler R TITLE Redundancy of human ATG4 protease isoforms in autophagy and LC3/GABARAP processing revealed in cells. JOURNAL Autophagy. 2019 Jan 21. doi: 10.1080/15548627.2019.1569925. PUBMED 30661429 REFERENCE 2 (bases 1 to 3144) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3144 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 94..111 /label=L4440 /note="L4440 vector, forward primer" terminator 270..297 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 389..475 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 524..546 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 538..555 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 539..555 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 571..670 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1025..1124) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1141..1160) /label=T7 /note="T7 promoter, forward primer" promoter complement(1142..1160) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1165..1181) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1165..1181) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1612..2403 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 2496..3084 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2985..3004 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc 241 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct 301 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct 361 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca 421 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc 481 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt 541 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg 601 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa 661 aaagcaggct taatgaagtt cgtgtacaaa gaagagcatc cgttcgagaa gcgccgctct 721 gagggcgaga agatccgaaa gaaatacccg gaccgggtgc cggtgatagt agaaaaggct 781 cccaaagctc ggataggaga cctggacaaa aagaaatacc tggtgccttc tgatctcaca 841 gttggtcagt tctacttctt gatccggaag cgaattcatc tccgagctga ggatgccttg 901 tttttctttg tcaacaatgt cattccaccc accagtgcca caatgggtca gctgtaccag 961 gaacaccatg aagaagactt ctttctctac attgcctaca gtgacgaaag tgtctacggt 1021 ctgtacccag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt 1081 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc 1141 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct 1201 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc 1261 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg 1321 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa 1381 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt 1441 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg 1501 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc 1561 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa 1621 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat 1681 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag 1741 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg 1801 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc 1861 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct 1921 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag 1981 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt 2041 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt 2101 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag 2161 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg 2221 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc 2281 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca 2341 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa 2401 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga 2461 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 2521 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 2581 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 2641 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 2701 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 2761 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 2821 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 2881 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 2941 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 3001 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 3061 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 3121 ttttgctggc cttttgctca catg //