LOCUS Exported 4838 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses CIRTS-10 (bdefensin 3-SLBP-YTHDF2(1-200)) in mammalian cells. ACCESSION . VERSION . KEYWORDS CIRTS-10: bdefensin 3-SLBP-YTHDF2(1-200) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4838) AUTHORS Rauch S, He E, Srienc M, Zhou H, Zhang Z, Dickinson BC TITLE Programmable RNA-Guided RNA Effector Proteins Built from Human Parts. JOURNAL Cell. 2019 Jun 27;178(1):122-134.e12. doi: 10.1016/j.cell.2019.05.049. Epub 2019 Jun 20. PUBMED 31230714 REFERENCE 2 (bases 1 to 4838) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4838 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 46..65 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(348..936) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(428..447) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 1052..1335 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers" enhancer 1526..1905 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1906..2109 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2060..2080 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" intron 2245..2377 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2360..2379 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 2421..2430 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2844..2870 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS 3504..3527 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(3645..3663) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 3700..3821 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3711..3730) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3765..3784 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS 4220..176 /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4274..4293) /label=Neo-R /note="Neomycin resistance gene, reverse primer" ORIGIN 1 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 61 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 121 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 181 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag 241 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 301 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 361 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 421 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 481 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 541 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 601 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 661 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 721 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 781 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 841 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 901 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg 961 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 1021 aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg 1081 cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta 1141 atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga 1201 ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc 1261 tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc 1321 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat 1381 cgagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat 1441 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc 1501 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca 1561 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta 1621 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat 1681 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg 1741 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac 1801 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt 1861 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg 1921 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc 1981 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt 2041 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata 2101 agcagagctg gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac 2161 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcaga 2221 agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac 2281 caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt 2341 ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac 2401 agctcttaag gctagagtat gccaccatgg gaattatcaa cacccttcaa aaatactact 2461 gtcgcgtccg tgggggtcgt tgcgcagtcc tgtcgtgcct gcctaaagag gagcagatcg 2521 gtaaatgctc aacccgtgga cgtaagtgct gtcgtcgcaa gaaaggcggt tctggaggct 2581 caggggggag cggagggtcc ggtggatcag gaggctccgc ggacttcgag acggatgaga 2641 gtgtactgat gcgccgccag aagcaaatca attacggaaa gaacactatc gcatacgacc 2701 gctatatcaa ggaagtgcca cgccacttgc gccagcccgg catccatcca aaaacgccca 2761 ataagtttaa aaagtattca cgtcgttcgt gggatcaaca aattaaactg tggaaagttg 2821 cacttcactt ctgggacctt caactgcctc cacttgaaag actgacactg ggcggtagtg 2881 gaggaagcgg ggggagcgga gggagtggtg gatccggcgg ctccatgtcg gccagcagcc 2941 tcttggagca gagaccaaaa ggtcaaggaa acaaagtaca aaatggatct gtacatcaaa 3001 aggatggatt aaacgatgat gattttgaac cttacttgag tccacaggca aggcccaata 3061 atgcatatac tgccatgtca gattcctact tacccagtta ctacagtccc tccattggct 3121 tctcctattc tttgggtgaa gctgcttggt ctacgggggg tgacacagcc atgccctact 3181 taacttctta tggacagctg agcaacggag agccccactt cctaccagat gcaatgtttg 3241 ggcaaccagg agccctaggt agcactccat ttcttggtca gcatggtttt aatttctttc 3301 ccagtgggat tgacttctca gcatggggaa ataacagttc tcagggacag tctactcaga 3361 gctctggata tagtagcaat tatgcttatg cacctagctc cttaggtgga gccatgattg 3421 atggacagtc agcttttgcc aatgagaccc tcaataaggc tcctggcatg aatactatag 3481 accaagggat ggcagcagga tccgattaca aggacgacga tgacaagtag tgtttaaacg 3541 aattcgggct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagctgatcc 3601 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact 3661 agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg 3721 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta 3781 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc 3841 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct 3901 acaaatgtgg taaaatcgaa ttctaatgga tcctctttgc gcttgcgttt tcccttgtcc 3961 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct 4021 acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag cgtgagcttc 4081 aaaagaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg 4141 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac 4201 aggatgacgg tcgtttcgca tgcttgaaca agatggattg cacgcaggtt ctccggccgc 4261 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 4321 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 4381 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 4441 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 4501 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 4561 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 4621 ccaccaagcg aaacatcgca tcgagcgagc acgcactcgg atggaagccg gtcttgtcga 4681 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 4741 caaggcgcgt atgccggatg gtgaggatct cgtcgtgact catggcgatg cctgcttgcc 4801 gaatatcatg gtggaaaatg gccgcttttc tggattca //