LOCUS Exported 5676 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-cdh1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5676) AUTHORS Pfleger CM, Lee E, Kirschner MW TITLE Substrate recognition by the Cdc20 and Cdh1 components of the anaphase-promoting complex. JOURNAL Genes Dev. 2001 Sep 15. 15(18):2396-407. PUBMED 11562349 REFERENCE 2 (bases 1 to 5676) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5676 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 7..984 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1019..1037 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1019..1036 /label=SP6 /note="SP6 promoter, forward primer" CDS 1110..1136 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1140..1166 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(1770..1781) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter complement(2690..2707) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2712..2846 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2736..2755) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2790..2809 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2959..2979) /label=T3 /note="T3 promoter, forward primer" promoter complement(2959..2977) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2998..3014) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2998..3014) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3011..3033) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3022..3038 /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(3046..3076) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3091..3112 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3229..3246) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3400..3988) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3480..3499) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4159..5019) /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 4782..4801 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5020..5124) /gene="bla" /label=AmpR promoter rep_origin complement(5150..5605) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5287..5308) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5499..5518 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg 61 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg 121 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg 181 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg 241 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg 301 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat 361 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt 421 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc 481 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct 541 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt 601 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga 661 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa 721 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta 781 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg 841 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg 901 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat 961 ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat 1021 ttaggtgaca ctatagaata caagctactt gttctttttg caggatccca tcggggatgg 1081 agctccaccg cggtggcggc cgcatctttt acccatacga tgttcctgac tatgcgggct 1141 atccctatga cgtcccggac tatgcaggat ccgattcgaa ttcgatggac caggactatg 1201 agcggcgcct gcttcgccag atcgtcatcc agaatgagaa cacgatgcca cgcgtcacag 1261 agatgcggcg gaccctgacg cctgccagct ccccagtgtc ctcgcccagc aagcacggag 1321 accgcttcat cccctccaga gccggagcca actggagcgt gaacttccac aggattaacg 1381 agaatgagaa gtctcccagt cagaaccgga aagccaagga cgccacctca gacaacggca 1441 aagacggcct ggcctactct gccctgctca agaatgagct gctgggtgcc ggcatcgaga 1501 aggtgcagga cccgcagact gaggaccgca gactgcagcc ctccacgcct gagaagaagg 1561 gtctgttcac gtattccctt agcaccaagc gctccagccc cgatgacggc aacgatgtgt 1621 ctccctactc cctgtctccc gtcagcaaca agagccagaa gctgctccgg tccccccgga 1681 aacccacccg caagatctcc aagatcccct tcaaggtgct ggacgcgccc gagctgcagg 1741 acgacttcta cctcaatctg gtggactggt cgtccctcaa tgtgctcagc gtggggctag 1801 gcacctgcgt gtacctgtgg agtgcctgta ccagccaggt gacgcggctc tgtgacctct 1861 cagtggaagg ggactcagtg acctccgtgg gctggtctga gcgggggaac ctggtggcgg 1921 tgggcacaca caagggcttc gtgcagatct gggacgcagc cgcagggaag aagctgtcca 1981 tgttggaggg ccacacggca cgcgtcgggg cgctggcctg gaatgctgag cagctgtcgt 2041 ccgggagccg cgaccgcatg atcctgcaga gggacatccg caccccgcca ctgcagtcgg 2101 agcggcggct gcagggccac cggcaggagg tgtgcgggct caagtggtcc acagaccacc 2161 agctcctcgc ctcggggggc aacgacaaca agctgctggt ctggaatcac tcgagcctga 2221 gccccgtgca gcagtacacg gagcacctgg cggccgtgaa ggccatcgcc tggtccccac 2281 atcagcacgg gctgctggcc tcggggggcg gcacagctga ccgctgtatc cgcttctgga 2341 acacgctgac aggacaacca ctgcagtgta tcgacacggg ctcccaagtg tgcaatctgg 2401 cctggtccaa gcacgccaac gagctggtga gcacgcacgg ctactcacag aaccagatcc 2461 ttgtctggaa gtacccctcc ctgacccagg tggccaagct gaccgggcac tcctaccgcg 2521 tgctgtacct ggcaatgtcc cctgatgggg aggccatcgt cactggtgct ggagacgaga 2581 ccctgaggtt ctggaacgtc tttagcaaaa cccgttcgac aaaggagtct gtgtctgtgc 2641 tcaacctctt caccaggatc cggtaatcta gcctctcgag cctctagaac tatagtgagt 2701 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta 2761 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 2821 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 2881 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc 2941 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc 3001 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 3061 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 3121 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3181 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 3241 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 3301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 3361 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 3421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 3481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 3541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 3601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 3661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 3721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 3781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 3841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 3901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 4081 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 4141 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 4201 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 4261 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 4321 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 4381 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 4441 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 4501 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 4561 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 4621 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 4681 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 4741 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 4801 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 4861 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 4921 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 4981 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 5041 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 5101 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc 5161 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 5221 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 5281 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 5341 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 5401 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 5461 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 5521 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 5581 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa 5641 gggcgatcgg tgcgggcctc ttcgctatta cgccag //