LOCUS Exported 6115 bp ds-DNA circular SYN 27-MAR-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6115) AUTHORS More TITLE Direct Submission JOURNAL Exported Monday, Mar 30, 2020 from SnapGene Viewer 5.0.4 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6115 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..19 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" /note="color: #ffffff; direction: RIGHT" 5'UTR 44..86 /label=Xenopus globin 5'-UTR /note="translational enhancer from the 5'-UTR of the major beta-globin gene of Xenopus laevis" /note="color: #a6acb3; direction: RIGHT" regulatory 102..111 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /note="color: #a6acb3" primer_bind 105..140 /label=13d_Hs_FW_Not /note="color: black; sequence: aaaaccatggcgagcgaggccagcatcgaaaaaaaaaag" misc_feature 108..3071 /label=HsCas13d /note="color: #993300" primer_bind complement(3014..3077) /label=13d_Hs_Rv_SacII /note="color: black; sequence: aaaaCCGCGGttaagcgtaatctggaacatcgtatgggtaagcggccgctccggatccg gaattgccg" CDS 3042..3068 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /note="color: #cc99b2" /translation="YPYDVPDYA" primer_bind complement(3046..3077) /label=13d_NLS_Hs_Rv_SacII /note="color: black; sequence: aaaaCCGCGGttaagcgtaatctggaacatcgtatg" 3'UTR 3093..3218 /label=Xenopus globin 3'-UTR /note="3'-UTR of the major beta-globin gene of Xenopus laevis" /note="color: #a6acb3; direction: RIGHT" promoter complement(3305..3323) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff; direction: LEFT" primer_bind complement(3330..3346) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" rep_origin complement(3487..3942) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" promoter 3969..4073 /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: RIGHT" CDS 4074..4934 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 4074 .. 4142 / #ccffcc / signal sequence 2: 4143 .. 4934 / #ccffcc Cleavage site after base 4142" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 5105..5693 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" protein_bind 5981..6002 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b" promoter 6017..6047 /label=lac promoter /note="promoter for the E. coli lac operon" /note="This forward directional feature has 3 segments: 1: 6017 .. 6022 / #ffffff / -35 2: 6023 .. 6040 / #ffffff 3: 6041 .. 6047 / #ffffff / -10" protein_bind 6055..6071 /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)." /note="color: #31849b" primer_bind 6079..6095 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: RIGHT" ORIGIN 1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga 61 agctcagaat aaacgctcaa ctttggcaga tctaactcga cgccaccatg gcgagcgagg 121 ccagcatcga aaaaaaaaag tccttcgcca agggcatggg cgtgaagtcc acactcgtgt 181 ccggctccaa agtgtacatg acaaccttcg ccgaaggcag cgacgccagg ctggaaaaga 241 tcgtggaggg cgacagcatc aggagcgtga atgagggcga ggccttcagc gctgaaatgg 301 ccgataaaaa cgccggctat aagatcggca acgccaaatt cagccatcct aagggctacg 361 ccgtggtggc taacaaccct ctgtatacag gacccgtcca gcaggatatg ctcggcctga 421 aggaaactct ggaaaagagg tacttcggcg agagcgctga tggcaatgac aatatttgta 481 tccaggtgat ccataacatc ctggacattg aaaaaatcct cgccgaatac attaccaacg 541 ccgcctacgc cgtcaacaat atctccggcc tggataagga cattattgga ttcggcaagt 601 tctccacagt gtatacctac gacgaattca aagaccccga gcaccatagg gccgctttca 661 acaataacga taagctcatc aacgccatca aggcccagta tgacgagttc gacaacttcc 721 tcgataaccc cagactcggc tatttcggcc aggccttttt cagcaaggag ggcagaaatt 781 acatcatcaa ttacggcaac gaatgctatg acattctggc cctcctgagc ggactgaggc 841 actgggtggt ccataacaac gaagaagagt ccaggatctc caggacctgg ctctacaacc 901 tcgataagaa cctcgacaac gaatacatct ccaccctcaa ctacctctac gacaggatca 961 ccaatgagct gaccaactcc ttctccaaga actccgccgc caacgtgaac tatattgccg 1021 aaactctggg aatcaaccct gccgaattcg ccgaacaata tttcagattc agcattatga 1081 aagagcagaa aaacctcgga ttcaatatca ccaagctcag ggaagtgatg ctggacagga 1141 aggatatgtc cgagatcagg aaaaatcata aggtgttcga ctccatcagg accaaggtct 1201 acaccatgat ggactttgtg atttataggt attacatcga agaggatgcc aaggtggctg 1261 ccgccaataa gtccctcccc gataatgaga agtccctgag cgagaaggat atctttgtga 1321 ttaacctgag gggctccttc aacgacgacc agaaggatgc cctctactac gatgaagcta 1381 atagaatttg gagaaagctc gaaaatatca tgcacaacat caaggaattt aggggaaaca 1441 agacaagaga gtataagaag aaggacgccc ctagactgcc cagaatcctg cccgctggcc 1501 gtgatgtttc cgccttcagc aaactcatgt atgccctgac catgttcctg gatggcaagg 1561 agatcaacga cctcctgacc accctgatta ataaattcga taacatccag agcttcctga 1621 aggtgatgcc tctcatcgga gtcaacgcta agttcgtgga ggaatacgcc tttttcaaag 1681 actccgccaa gatcgccgat gagctgaggc tgatcaagtc cttcgctaga atgggagaac 1741 ctattgccga tgccaggagg gccatgtata tcgacgccat ccgtatttta ggaaccaacc 1801 tgtcctatga tgagctcaag gccctcgccg acaccttttc cctggacgag aacggaaaca 1861 agctcaagaa aggcaagcac ggcatgagaa atttcattat taataacgtg atcagcaata 1921 aaaggttcca ctacctgatc agatacggtg atcctgccca cctccatgag atcgccaaaa 1981 acgaggccgt ggtgaagttc gtgctcggca ggatcgctga catccagaaa aaacagggcc 2041 agaacggcaa gaaccagatc gacaggtact acgaaacttg tatcggaaag gataagggca 2101 agagcgtgag cgaaaaggtg gacgctctca caaagatcat caccggaatg aactacgacc 2161 aattcgacaa gaaaaggagc gtcattgagg acaccggcag ggaaaacgcc gagagggaga 2221 agtttaaaaa gatcatcagc ctgtacctca ccgtgatcta ccacatcctc aagaatattg 2281 tcaatatcaa cgccaggtac gtcatcggat tccattgcgt cgagcgtgat gctcaactgt 2341 acaaggagaa aggctacgac atcaatctca agaaactgga agagaaggga ttcagctccg 2401 tcaccaagct ctgcgctggc attgatgaaa ctgcccccga taagagaaag gacgtggaaa 2461 aggagatggc tgaaagagcc aaggagagca ttgacagcct cgagagcgcc aaccccaagc 2521 tgtatgccaa ttacatcaaa tacagcgacg agaagaaagc cgaggagttc accaggcaga 2581 ttaacaggga gaaggccaaa accgccctga acgcctacct gaggaacacc aagtggaatg 2641 tgatcatcag ggaggacctc ctgagaattg acaacaagac atgtaccctg ttcagaaaca 2701 aggccgtcca cctggaagtg gccaggtatg tccacgccta tatcaacgac attgccgagg 2761 tcaattccta cttccaactg taccattaca tcatgcagag aattatcatg aatgagaggt 2821 acgagaaaag cagcggaaag gtgtccgagt acttcgacgc tgtgaatgac gagaagaagt 2881 acaacgatag gctcctgaaa ctgctgtgtg tgcctttcgg ctactgtatc cccaggttta 2941 agaacctgag catcgaggcc ctgttcgata ggaacgaggc cgccaagttc gacaaggaga 3001 aaaagaaggt gtccggcaat tccggatccg gagcggccgc ttacccatac gatgttccag 3061 attacgctta accgcggatc tggttaccac taaaccagcc tcaagaacac ccgaatggag 3121 tctctaagct acataatacc aacttacact ttacaaaatg ttgtccccca aaatgtagcc 3181 attcgtatct gctcctaata aaaagaaagt ttcttcacat tctaaaaaaa aaaaaaaaaa 3241 aaaaaaaaaa aacccccccc cccccctgca gggccaagtc ggcctctaga ggtacccaat 3301 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg 3361 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg 3421 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc 3481 gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 3541 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 3601 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 3661 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 3721 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 3781 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 3841 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 3901 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg 3961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 4021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 4081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 4141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 4201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 4261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 4321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 4381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 4441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 4501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 4561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 4621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 4681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 4741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 4801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 4861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 4921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 4981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 5041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 5101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 5161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 5221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 5281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 5341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 5401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 5461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 5521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 5581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 5641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 5701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 5761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 5821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 5881 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca 5941 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc 6001 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga 6061 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctcga //