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" enhancer 93..472 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 473..676 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 627..647 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" intron 812..944 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 927..946 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 988..997 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1411..1437 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS 2071..2094 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(2212..2230) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 2267..2388 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2278..2297) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS 2787..3581 /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(2841..2860) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3451..3470 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(3753..4341) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3833..3852) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 4457..4740 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers" ORIGIN 1 aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta 61 tattggctca tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata 121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc 361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg 421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 481 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 601 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt 661 ctatataagc agagctggtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt 721 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag 781 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta 841 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc 901 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt 961 caattacagc tcttaaggct agagtatgcc accatgggaa ttatcaacac ccttcaaaaa 1021 tactactgtc gcgtccgtgg gggtcgttgc gcagtcctgt cgtgcctgcc taaagaggag 1081 cagatcggta aatgctcaac ccgtggacgt aagtgctgtc gtcgcaagaa aggcggttct 1141 ggaggctcag gggggagcgg agggtccggt ggatcaggag gctccgcgga cttcgagacg 1201 gatgagagtg tactgatgcg ccgccagaag caaatcaatt acggaaagaa cactatcgca 1261 tacgaccgct atatcaagga agtgccacgc cacttgcgcc agcccggcat ccatccaaaa 1321 acgcccaata agtttaaaaa gtattcacgt cgttcgtggg atcaacaaat taaactgtgg 1381 aaagttgcac ttcacttctg ggaccttcaa ctgcctccac ttgaaagact gacactgggc 1441 ggtagtggag gaagcggggg gagcggaggg agtggtggat ccggcggctc catgtcggcc 1501 agcagcctct tggagcagag accaaaaggt caaggaaaca aagtacaaaa tggatctgta 1561 catcaaaagg atggattaaa cgatgatgat tttgaacctt acttgagtcc acaggcaagg 1621 cccaataatg catatactgc catgtcagat tcctacttac ccagttacta cagtccctcc 1681 attggcttct cctattcttt gggtgaagct gcttggtcta cggggggtga cacagccatg 1741 ccctacttaa cttcttatgg acagctgagc aacggagagc cccacttcct accagatgca 1801 atgtttgggc aaccaggagc cctaggtagc actccatttc ttggtcagca tggttttaat 1861 ttctttccca gtgggattga cttctcagca tggggaaata acagttctca gggacagtct 1921 actcagagct ctggatatag tagcaattat gcttatgcac ctagctcctt aggtggagcc 1981 atgattgatg gacagtcagc ttttgccaat gagaccctca ataaggctcc tggcatgaat 2041 actatagacc aagggatggc agcaggatcc gattacaagg acgacgatga caagtagtgt 2101 ttaaacgaat tcgggctcgg tacacgggga tcctctagag tcgacctgca ggcatgcaag 2161 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc 2221 aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat 2281 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt 2341 gatgctattt ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat 2401 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa 2461 aacctctaca aatgtggtaa aatcgaattc taatggatcc tctttgcgct tgcgttttcc 2521 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg 2581 gctttctacg tgttccgctt cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt 2641 gagcttcaaa agaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag 2701 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc 2761 aagagacagg atgacggtcg tttcgcatgc ttgaacaaga tggattgcac gcaggttctc 2821 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 2881 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 2941 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 3001 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3061 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3121 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 3181 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg cactcggatg gaagccggtc 3241 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 3301 ccaggctcaa ggcgcgtatg ccggatggtg aggatctcgt cgtgactcat ggcgatgcct 3361 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 3421 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 3481 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 3541 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 3601 aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt aatacggtta 3661 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 3721 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 3781 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 3841 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 3901 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 3961 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 4021 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 4081 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 4141 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 4201 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 4261 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 4321 cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc tgacgctcag 4381 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 4441 tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt cgctgggatg 4501 gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg gcctgtaccc 4561 tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg gcagtttttc 4621 gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac gcgttttagc 4681 ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga 4741 gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga tcttcaatat 4801 tggccattag ccatattatt cattggttat atagcata //