LOCUS Exported 6059 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.2 V5-DEST 3XFlag-SRSF3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6059) 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 6059) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6059 /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" CDS 939..1004 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" protein_bind complement(1499..1523) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1549..1590 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(1630..1649) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1674..1722 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 1924..2352 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2011..2030) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2221..2242 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2361..2381) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2366..2695 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2546..2681 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2608..2627 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2762..3556 /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(2816..2835) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3426..3445 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3732..3853 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3769..3788) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3823..3842 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3902..3918) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3902..3918) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3915..3937) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3926..3942 /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(3950..3980) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3995..4016 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4133..4150) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4304..4892) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4384..4403) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5063..5923) /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 5686..5705 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5924..6028) /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 aggctatgga ctacaaagac catgacggtg 961 attataaaga tcatgatatc gattacaagg atgacgatga caagggccat cgtgattcct 1021 gtccattgga ctgtaaggtt tatgtaggca atcttggaaa caatggcaac aagacggaat 1081 tggaacgggc ttttggctac tatggaccac tccgaagtgt gtgggttgct agaaacccac 1141 ccggctttgc ttttgttgaa tttgaagatc cccgagatgc agctgatgca gtccgagagc 1201 tagatggaag aacactatgt ggctgccgtg taagagtgga actgtcgaat ggtgaaaaaa 1261 gaagtagaaa tcgtggccca cctccctctt ggggtcgtcg ccctcgagat gattatcgta 1321 ggaggagtcc tccacctcgt cgcagatctc caagaaggag aagcttctct cgcagccgga 1381 gcaggtccct ttctagagat aggagaagag agagatcgct gtctcgggag agaaatcaca 1441 agccgtcccg atccttctct aggtctcgta gtcgatctag gtcaaatgaa aggaaatagc 1501 ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg taagcctatc 1561 cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgagt ttaaacgggg 1621 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 1681 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt 1741 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg 1801 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca 1861 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc 1921 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 1981 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 2041 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat 2101 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 2161 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 2221 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 2281 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 2341 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 2401 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 2461 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 2521 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 2581 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc 2641 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 2701 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg 2761 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 2821 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 2881 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 2941 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 3001 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 3061 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 3121 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 3181 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 3241 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 3301 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 3361 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 3421 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 3481 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 3541 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca 3601 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 3661 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 3721 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 3781 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 3841 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 3901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 3961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat 4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 4741 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 4921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 4981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 5101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 5161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 5221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 5281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 5341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 5401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 5461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 5521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 5581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 5641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 5701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 5761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 5821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 5881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 5941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 6001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //