LOCUS Exported 6961 bp ds-DNA circular SYN 25-JAN-2022 DEFINITION encodes multiple copies of an epitope tag HA. ACCESSION . VERSION . KEYWORDS pCAG_smFP HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6961) AUTHORS Viswanathan S, Williams ME, Bloss EB, Stasevich TJ, Speer CM, Nern A, Pfeiffer BD, Hooks BM, Li WP, English BP, Tian T, Henry GL, Macklin JJ, Patel R, Gerfen CR, Zhuang X, Wang Y, Rubin GM, Looger LL TITLE High-performance probes for light and electron microscopy. JOURNAL Nat Methods. 2015 Jun;12(6):568-76. doi: 10.1038/nmeth.3365. Epub 2015 Apr 27. PUBMED 25915120 REFERENCE 2 (bases 1 to 6961) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 25, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6961 /organism="synthetic DNA construct" /mol_type="other DNA" intron 122..1138 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1062..1084 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1146..1165 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1264..1290 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1294..1320 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1324..1350 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1879..1905 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1912..1938 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1951..1977 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1984..2010 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2221..2247 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2251..2277 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2281..2307 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" intron 2356..2488 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2471..2490 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 2497..3085 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2550..2570) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2968..2979) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3112..3129) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3118..3229 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3325..3345) /label=T3 /note="T3 promoter, forward primer" promoter complement(3325..3343) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3364..3380) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3482..3501) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3547..3602 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3601..3620) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3963..3979) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3987..4003 /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(4011..4041) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4056..4077 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4136..4331 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4182..4317 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4244..4263 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4337..4471 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4374..4393) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4428..4447 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4539..4556) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4710..5298) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4790..4809) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5469..6329) /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 6092..6111 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6330..6434) /gene="bla" /label=AmpR promoter enhancer 6465..6844 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 61 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 121 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc 181 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc 241 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga 301 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg 361 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg 421 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg 481 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt 541 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca 601 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg 661 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc 721 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg 781 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc 841 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac 901 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga 961 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc 1021 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt 1081 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct 1141 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccgc 1201 gggcccggcg cgccagctgg atatctccgg acccgggcta gcgcaattgc ggccgccacc 1261 atgtaccctt atgatgtgcc cgattatgct ggctaccctt atgatgttcc tgattacgcc 1321 ggatatccgt atgacgtgcc agactacgcg ggaggtgtga gcaagggaga ggagttgttt 1381 accggcgtgg tccccatact ggtggagttg gatggcgacg ttaatggaca taaattctcg 1441 gtccgcggcg agggagaggg agacgccacc aatggcaagc tgacgcttaa gttcatttgc 1501 acgactggaa aattgcctgt cccctggccg accctggtca ccacactggg cggaggagtg 1561 cagtgcttct cccgttaccc agaccacatg aagcagcacg atttcttcaa gagcgcaatg 1621 ccggagggat acgtgcaaga acggactatt tccttcaagg atgacggaac ctataagacc 1681 cgtgcggagg tcaaattcga gggtgacacc ctggtgaacc gaattgaact caaaggaatc 1741 gatttcaagg aggatggaaa tattctgggt cacaagctgg aatacaactt caacagccat 1801 aatgtgtaca ttacggctga taagcagaag aacggcatta aggccaattt caagatccgc 1861 cacaacgttg agggtggata tccctacgac gtgcccgatt atgcgggcgg atatccgtat 1921 gatgttccag attatgctgg tggtggaggc tatccctatg atgtccccga ctacgccgga 1981 ggttacccat acgacgtgcc cgactacgct ggaggcgatg gcagcgtgca gctggcagat 2041 cattatcaac agaatacccc cataggtgat ggccccgttc tgcttccaga taatcactac 2101 ctttccaccc agagcgtgct ttcgaaagac ccgaacgaaa agcgtgatca catggtcctg 2161 ctggagtttg tgaccgcggc tggaatcacc ctgggtatgg acgaactcta caagggtggc 2221 tacccctacg atgtgccgga ttacgctgga tatccgtatg acgtaccgga ctatgccggt 2281 tacccgtacg atgtcccgga ctacgcttaa accggtgctc gaggtaccga tgaatagcta 2341 aggtcgaggc cgcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa 2401 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact 2461 gacatccact ttgcctttct ctccacaggt gtcgacaatc aacctctgga ttacaaaatt 2521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 2581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 2641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 2701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 2761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 2821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 2881 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 2941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 3001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 3061 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtacgatca gcctcgactg 3121 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 3181 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 3241 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 3301 aagacaatag cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc 3361 atggtcatag ctgtttcctg tgtgaaattg ttatccgcta attcactcct caggtgcagg 3421 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag 3481 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt 3541 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct 3601 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt 3661 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag 3721 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga 3781 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa 3841 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca 3901 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca 3961 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4021 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc 4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4261 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4321 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4441 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg 4501 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 4561 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 4621 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 4921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 5401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 5461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 5521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5581 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgacatt gattattgac 6481 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 6541 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 6601 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 6661 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 6721 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 6781 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 6841 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc 6901 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg 6961 g //