LOCUS Exported 5763 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA6.2-EmGFP-mir9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5763) AUTHORS Lau P, Verrier JD, Nielsen JA, Johnson KR, Notterpek L, Hudson LD TITLE Identification of dynamically regulated microRNA and mRNA networks in developing oligodendrocytes. JOURNAL J Neurosci. 2008 Nov 5. 28(45):11720-30. PUBMED 18987208 REFERENCE 2 (bases 1 to 5763) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5763 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 384..587 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 538..558 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 632..650 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 680..704 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 713..1432 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(758..779) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1019..1038) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1366..1387 /label=EGFP-C /note="EGFP, forward primer" ncRNA 1492..1518 /label=5' miR-155 /note="sequence upstream of the precursor of mouse miR-155 microRNA (Uva et al., 2013)" /ncRNA_class="miRNA" ncRNA 1586..1627 /label=3' miR-155 /note="sequence downstream of the precursor of mouse miR-155 microRNA (Uva et al., 2013)" /ncRNA_class="miRNA" protein_bind complement(1656..1680) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(1722..1741) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1766..1814 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" promoter complement(1985..2003) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2008..2024) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2008..2024) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" rep_origin 2092..2520 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2179..2198) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2389..2410 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2529..2549) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2534..2863 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2714..2849 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2776..2795 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 2911..2958 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 2977..3375 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 3533..3654 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3570..3589) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3624..3643 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3689..3706) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3860..4448) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3940..3959) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4544..5335) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" ORIGIN 1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagtcccaag 661 ctggctagtt aagctatcaa caagtttgta caaaaaagca ggctttaaaa ccatggtgag 721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 901 caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga 961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 1021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 1081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 1141 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa 1201 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 1261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 1321 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 1381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagctaagca 1441 cttcgtggcc gtcgatcgtt taaagggagg tagtgagtcg accagtggat cctggaggct 1501 tgctgaaggc tgtatgctgt ctttggttat ctagctgtat gagttttggc cactgactga 1561 ctcatacaga gataaccaaa gacaggacac aaggcctgtt actagcactc acatggaaca 1621 aatggcccag atctggccgc actcgagata tctagaccca gctttcttgt acaaagtggt 1681 tgatctagag ggcccgcggt tcgctgatgg gggaggctaa ctgaaacacg gaaggagaca 1741 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt 1801 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc 1861 accgtgaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc 1921 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc 1981 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg 2041 tctcaaaatc tctgatggat ctgcgcagct ggggctctag ggggtatccc cacgcgccct 2101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 2161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 2221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 2281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 2341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 2401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 2461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 2521 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 2881 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc 2941 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa 3001 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac 3061 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta 3121 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg 3181 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc 3241 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg 3301 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat 3361 gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt 3421 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 3481 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 3541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 3601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 3661 ctgtataccg tcgctcttcc gctgcttcct cgctcactga ctcgctgcgc tcggtcgttc 3721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 3781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 3841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 3901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 3961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 4021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 4081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 4141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 4201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 4261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 4321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 4381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 4441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgt 4501 aactcacgtt aagggatttt ggtcatgggt ggctcgacga gggttatttg ccgactacct 4561 tggtgatctc gcctttcacg tagtggacaa attcttccaa ctgatctgcg cgcgaggcca 4621 agcgatcttc ttcttgtcca agataagcct gtctagcttc aagtatgacg ggctgatact 4681 gggccggcag gcgctccatt gcccagtcgg cagcgacatc cttcggcgcg attttgccgg 4741 ttactgcgct gtaccaaatg cgggacaacg taagcactac atttcgctca tcgccagccc 4801 agtcgggcgg cgagttccat agcgttaagg tttcatttag cgcctcaaat agatcctgtt 4861 caggaaccgg atcaaagagt tcctccgccg ctggacctac caaggcaacg ctatgttctc 4921 ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt ggctggctcg aagatacctg 4981 caagaatgtc attgcgctgc cattctccaa attgcagttc gcgcttagct ggataacgcc 5041 acggaatgat gtcgtcgtgc acaacaatgg tgacttctac agcgcggaga atctcgctct 5101 ctccagggga agccgaagtt tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat 5161 caagccttac ggtcaccgta accagcaaat caatatcact gtgtggcttc aggccgccat 5221 ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga 5281 cgccaactac ctctgatagt tgagtcgata cttcggcgat caccgcttcc ctcataatgt 5341 ttaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctc cataacatca 5401 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtaccc 5461 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt 5521 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agcttacgaa 5581 ccgaacaggc ttatgtccac tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc 5641 cggcaacctt gggtagcagc gaagtcgagg catttctgtc ctggctggtc tagaattgca 5701 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata 5761 cgc //