LOCUS Exported 5265 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION 3'UTR segments of target genes can be inserted into this firefly luciferase reporter to test for their effects on protein production. ACCESSION . VERSION . KEYWORDS pIS0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5265) AUTHORS Yekta S, Shih IH, Bartel DP TITLE MicroRNA-directed cleavage of HOXB8 mRNA. JOURNAL Science. 2004 Apr 23. 304(5670):594-6. PUBMED 15105502 REFERENCE 2 (bases 1 to 5265) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5265 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 17..213 /label=SV40 promoter /note="SV40 early promoter" rep_origin 64..199 /label=SV40 ori /note="SV40 origin of replication" primer_bind 126..145 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" regulatory 243..252 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 249..1901 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(315..333) /label=LucNrev /note="Firefly luciferase, reverse primer" polyA_signal 1982..2103 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1993..2012) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2047..2066 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2425..2445 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" primer_bind complement(2597..2614) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2768..3356) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2848..2867) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3527..4387) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4150..4169 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4388..4492) /gene="bla" /label=AmpR promoter rep_origin 4519..4974 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4606..4625) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4816..4837 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 5105..5153 /note="synthetic polyadenylation signal" misc_feature 5167..5258 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 5207..5226 /label=RVprimer3 /note="pGL3 vector, forward primer" ORIGIN 1 gtcgagatct gcgatctgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 61 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg ctgactaatt 121 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga 181 ggaggctttt ttggaggcct aggcttttgc aaaaagcttg gcattccggt actgttggta 241 aagccaccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg 301 aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg 361 gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg 421 aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa 481 tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta 541 tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta 601 tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga 661 acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct aaaacggatt 721 accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg 781 aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact 841 cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga 901 gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt 961 taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat 1021 gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc 1081 aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa 1141 gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc 1201 ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc 1261 aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag ggggatgata 1321 aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata 1381 ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta 1441 tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc 1501 tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc 1561 tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct 1621 tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg 1681 gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga 1741 tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt 1801 ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga 1861 tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta attctaggag ctctatacgc 1921 gtctcaagct tactgctagc gttctagagt cggggcggcc ggccgcttcg agcagacatg 1981 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt 2041 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa 2101 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt 2161 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct gaacgatgga 2221 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga 2281 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg 2341 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg 2401 ctggggagcc tggggacttt ccacacccta actgacacac attccacagc ggatccgtcg 2461 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac 2521 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc 2581 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 2641 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 2701 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 2761 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 2821 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 2881 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 2941 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 3001 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 3061 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 3121 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 3181 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 3241 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 3301 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 3361 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 3421 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 3481 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 3541 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 3601 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 3661 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 3721 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 3781 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 3841 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 3901 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 3961 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 4021 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 4081 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 4141 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 4201 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 4261 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 4321 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 4381 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 4441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 4501 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 4561 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 4621 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 4681 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 4741 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 4801 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 4861 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 4921 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt 4981 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac 5041 gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg 5101 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat 5161 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc 5221 tgtccccagt gcaagtgcag gtgccagaac atttctctat cgata //