LOCUS Exported 3281 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION tTAF2 ORF with stop codon between attL1-attL2 on Gateway entry vector. ACCESSION . VERSION . KEYWORDS pENTR-tTAF2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3281) TITLE Minoru Ko Lab plasmids REFERENCE 2 (bases 1 to 3281) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3281 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 671..1288 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG" primer_bind complement(744..763) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" protein_bind complement(1406..1505) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1522..1541) /label=T7 /note="T7 promoter, forward primer" promoter complement(1523..1541) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1546..1562) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1546..1562) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1664..1683) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1675..2484 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1742..1761) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2631..3219 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3120..3139 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctcc atgtctcgat tagataaaag taaagtgatt aacagcgcat tagagctgct 721 taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga 781 gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat 841 tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga 901 ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc 961 aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt 1021 agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt 1081 ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga 1141 aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt 1201 tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt 1261 agaaaaacaa cttaaatgtg aaagtgggtc cccggccgac gccctggacg acttcgacct 1321 ggacatgctg cctgctgatg ctctcgatga tttcgatctg gatatgctcc cgggtaacta 1381 agtaaggatc tactggttct ggagcaccca gctttcttgt acaaagttgg cattataaga 1441 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt 1501 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc 1561 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata 1621 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc 1681 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat 1741 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat 1801 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat 1861 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc 1921 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa 1981 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg 2041 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat 2101 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt 2161 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa 2221 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt 2281 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac 2341 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag 2401 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat 2461 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc 2521 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac 2581 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 2641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 2701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 2761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 2821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 2881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 2941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 3001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 3061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 3121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 3181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 3241 ttacggttcc tggccttttg ctggcctttt gctcacatgt t //