LOCUS Exported 4310 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Entry vector with human U6 promoter driving mouse G alpha 12 miR30-based shRNA.. ACCESSION . VERSION . KEYWORDS pEN_hU6miR-G12-E SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4310) AUTHORS Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, Rebres R, Roach T, Seaman W, Simon MI, Fraser ID TITLE A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression. JOURNAL Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64. PUBMED 16945906 REFERENCE 2 (bases 1 to 4310) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4310 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 25..48 /label=pENTR-F /note="pENTR vectors, forward primer" protein_bind 67..166 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 426..667 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 426..446 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 598..617 /label=LKO.1 5' /note="Human U6 promoter, forward primer" ncRNA 730..851 /label=5' miR-30a /note="sequence upstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 879..893 /label=miR-30a loop /note="loop from the the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 921..1047 /label=3' miR-30a /note="sequence downstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" protein_bind complement(1080..1179) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1291..1310) /label=pENTR-R /note="pENTR vectors, reverse primer" primer_bind complement(1369..1388) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" CDS complement(1962..2495) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(2684..2712) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" rep_origin 3369..3957 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3858..3877 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 4122..4208 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 4300..17 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" ORIGIN 1 ctgacggatg gcctttttgc gtttctacaa actcttcctg ttagttagtt acttaagctc 61 gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat 121 aagcaatgct tttttataat gccaactttg tacaaaaaag caggctttaa aggaaccaat 181 tcagtcgact ggatcccccc gagtccaaca cccgtgggaa tcccatgggc accatggccc 241 ctcgctccaa aaatgctttc gcgtctcgca gacactgctc ggtagtttcg gggatcagcg 301 tttgagtaag agcccgcgtc tgaaccctcc gcgccgcccc ggcccagtgg aaagacgcgc 361 aggcaaaacg caccacgtga cggagcgtga ccgcgcgccg agcgcgcgcc aaggtcgggc 421 aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta 481 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac 541 gtagaaagta ataatttctt gggtagtttg cagtttttaa aattatgttt taaaatggac 601 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg 661 aaaggacgaa acaccgtgct cgcttcggca gcacatatac tagtcgacta gggataacag 721 ggtaattgtt tgaatgaggc ttcagtactt tacagaatcg ttgcctgcac atcttggaaa 781 cacttgctgg gattacttct tcaggttaac ccaacagaag gctcgagaag gtatattgct 841 gttgacagtg agcgacgaca ccatcttcga caacattagt gaagccacag atgtaatgtt 901 gtcgaagatg gtgtcgctgc ctactgcctc ggaattcaag gggctacttt aggagcaatt 961 atcttgttta ctaaaactga ataccttgct atctctttga tacattttta caaagctgaa 1021 ttaaaatggt ataaattaaa tcactttttt caattggcgg ccgcactcga gatatctaga 1081 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa 1141 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg cagctctggc 1201 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa 1261 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg 1321 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 1381 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg 1441 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 1501 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 1561 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg 1621 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgtccctgc 1681 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 1741 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 1801 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct 1861 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 1921 ggaaattagg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg 1981 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg 2041 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca 2101 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact 2161 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc 2221 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta 2281 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct 2341 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg 2401 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg 2461 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca 2521 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa 2581 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa 2641 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca 2701 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac 2761 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc 2821 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg 2881 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt 2941 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt 3001 gttcgcccag gactctagct atagttctag tggttggcta ctaataggtt gtattgatgt 3061 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg 3121 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga 3181 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa 3241 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc 3301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 3361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 3421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 3481 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 3541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 3601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 3661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 3721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 3781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 3841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 3901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 3961 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 4021 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgctagcat ggatctcggg 4081 gacgtctaac tactaagcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc 4141 agtcggaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta 4201 ggacaaatcc gccgggagcg gatttgaacg ttgtgaagca acggcccgga gggtggcggg 4261 caggacgccc gccataaact gccaggcatc aaactaagca gaaggccatc //