LOCUS Exported 6223 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.2 mir-1-1 reporter cel-mir-124 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6223) AUTHORS Auyeung VC, Ulitsky I, McGeary SE, Bartel DP TITLE Beyond Secondary Structure: Primary-Sequence Determinants License Pri-miRNA Hairpins for Processing. JOURNAL Cell. 2013 Feb 14;152(4):844-58. doi: 10.1016/j.cell.2013.01.031. PUBMED 23415231 REFERENCE 2 (bases 1 to 6223) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6223 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 911..935 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1336..1360) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1713..1754 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(1794..1813) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1838..1886 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2088..2516 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2175..2194) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2385..2406 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2525..2545) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2530..2859 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2710..2845 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2772..2791 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2926..3720 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2980..2999) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3590..3609 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3896..4017 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3933..3952) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3987..4006 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4066..4082) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4066..4082) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4079..4101) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4090..4106 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4114..4144) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4159..4180 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4297..4314) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4468..5056) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4548..4567) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5227..6087) /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 5850..5869 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6088..6192) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagctatca acaagtttgt acaaaaaagc aggcttcatt gattcacatc cattttgaga 961 aggatggttg ctactgctgc ttgatctcaa ggactgtgtg ctcccacttt tactcaaagt 1021 attttatttc ttcttcaagt gctctcttct ccatttttga atttccttcg caattgatac 1081 accatgtccc acttgtcatc tggcatgcac cctagtgact ttagtggaca tctaagtctt 1141 ccaactaagg cacgcggtga atgccacgtg gccatgatgg ggctcccatt gaatttgaaa 1201 ggttggttga tctatctatc tatctcttca tagttattca tccatatcgg tgtcacagaa 1261 tgaaagcgag agagctgact aattcatgta catttgatgc tctttccctc ccattttgcg 1321 tccattcatt aggataccca gctttcttgt acaaagtggt tgatctagag ggcccgcagt 1381 ggggtcagct tctaccgggg cggcgtcccg gggtcttgga actgcatgca gactgcctgc 1441 ttgggaaaca tacttcttta tatgcccata tggacctgct aagctatgga atgtaaagaa 1501 gtatgtatct caggccggga cctctctcgc cgcactgagg ggcactccac accacggggg 1561 ccgccgcggc cgacagccag ccaggcggct gtggtggggg tgaggctggg gcggcaggct 1621 gcctgggcgt tggtggacgg tcgtgtcccc gccagcattg ttctgtcact ttgcctgtgt 1681 cagcaggtgg aagtgctcac cagacattcg aaggtaagcc tatccctaac cctctcctcg 1741 gtctcgattc tacgcgtacc ggttagtaat gagtttaaac gggggaggct aactgaaaca 1801 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 1861 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc 1921 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc 1981 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg 2041 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag 2101 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 2161 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 2221 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca 2281 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 2341 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 2401 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg 2461 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 2521 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 2581 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 2641 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 2701 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 2761 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 2821 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 2881 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 2941 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3001 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3061 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 3121 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3181 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3241 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3301 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3361 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3421 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 3481 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3541 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 3601 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 3661 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 3721 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 3781 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 3841 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt 3901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 3961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 4021 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 4081 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4141 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4201 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 4261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 4321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 4381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 4441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 4501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 4561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 4621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 4681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 4741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 4801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 4861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 4921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 4981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 5041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 5581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 5641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 5701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 5761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 5821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 5881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 5941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6181 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //