LOCUS Exported 6185 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.2 mir-1-1 reporter cel-lin-4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6185) 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 6185) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6185 /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(1298..1322) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1675..1716 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(1756..1775) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1800..1848 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 2050..2478 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2137..2156) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2347..2368 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2487..2507) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2492..2821 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2672..2807 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2734..2753 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2888..3682 /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(2942..2961) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3552..3571 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3858..3979 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3895..3914) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3949..3968 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4028..4044) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4028..4044) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4041..4063) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4052..4068 /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(4076..4106) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4121..4142 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4259..4276) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4430..5018) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4510..4529) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5189..6049) /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 5812..5831 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6050..6154) /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 aggctcatat gctaccacaa tcggtcggac 961 tcatcacact tacctttcaa atatctaaac tattcctgaa tataataaat cttatagttt 1021 ttagattcta gacaatttct agagttttgg ttggtttatg agtttatgct tccggcctgt 1081 tccctgagac ctcaagtgtg agtgtactat tgatgcttca cacctgggct ctccgggtac 1141 caggacggtt tgagcagatc tttttttctg ttttcacggg gttttttttc acactgggag 1201 ctagttccaa taaaatggta ttcggggaaa agggtcaaac gattattttc agaagtcgac 1261 tacaatagag ttccactaac gccaactgat gaagcttacc cagctttctt gtacaaagtg 1321 gttgatctag agggcccgca gtggggtcag cttctaccgg ggcggcgtcc cggggtcttg 1381 gaactgcatg cagactgcct gcttgggaaa catacttctt tatatgccca tatggacctg 1441 ctaagctatg gaatgtaaag aagtatgtat ctcaggccgg gacctctctc gccgcactga 1501 ggggcactcc acaccacggg ggccgccgcg gccgacagcc agccaggcgg ctgtggtggg 1561 ggtgaggctg gggcggcagg ctgcctgggc gttggtggac ggtcgtgtcc ccgccagcat 1621 tgttctgtca ctttgcctgt gtcagcaggt ggaagtgctc accagacatt cgaaggtaag 1681 cctatcccta accctctcct cggtctcgat tctacgcgta ccggttagta atgagtttaa 1741 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac 1801 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg 1861 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 1921 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 1981 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg 2041 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 2101 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 2161 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt 2221 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 2281 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 2341 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 2401 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac 2461 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 2521 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 2581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 2641 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 2701 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 2761 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 2821 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc 2881 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 2941 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 3001 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 3061 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 3121 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 3181 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 3241 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 3301 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 3361 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat 3421 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 3481 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 3541 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 3601 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 3661 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga 3721 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 3781 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 3841 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 3901 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 3961 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 4021 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 4081 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 4141 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 4201 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 4261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4621 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4861 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 4981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 5041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 5101 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 5161 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 5221 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 5281 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 5341 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 5401 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 5461 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 5521 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 5581 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 5641 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 5701 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 5761 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 5821 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 5881 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 5941 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 6001 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 6061 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 6121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 6181 acgtc //