LOCUS Exported 10143 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses dog ACE2 in mammalian cells. ACCESSION . VERSION . KEYWORDS pscALPSpuro-ClACE2 (Dog) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10143) AUTHORS Yurkovetskiy L, Wang X, Pascal KE, Tomkins-Tinch C, Nyalile TP, Wang Y, Baum A, Diehl WE, Dauphin A, Carbone C, Veinotte K, Egri SB, Schaffner SF, Lemieux JE, Munro JB, Rafique A, Barve A, Sabeti PC, Kyratsous CA, Dudkina NV, Shen K, Luban J TITLE Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant. JOURNAL Cell. 2020 Sep 15. pii: S0092-8674(20)31229-0. doi: 10.1016/j.cell.2020.09.032. PUBMED 32991842 REFERENCE 2 (bases 1 to 10143) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10143 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(64..84) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 69..398 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 249..384 /label=SV40 ori /note="SV40 origin of replication" primer_bind 311..330 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" enhancer 410..789 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 791..989 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 944..964 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 1007..1187 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1234..1359 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1856..2089 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2273..2317 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2615..2732 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2869..3276 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" CDS 5716..5745 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 6249..6848 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6249..6268) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6745..6765 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 6858..7446 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6911..6931) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7329..7340) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7964..8144 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(8215..8803) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8295..8314) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8974..9834) /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 9597..9616 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9835..9939) /gene="bla" /label=AmpR promoter primer_bind complement(10014..10033) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc 61 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 121 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 241 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 301 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 361 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg acattgatta 421 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 481 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 541 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 601 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 661 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 721 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 781 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 841 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 901 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 961 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt 1021 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 1081 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 1141 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac 1201 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct 1261 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact 1321 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt 1381 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa 1441 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa 1501 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca 1561 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata 1621 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag 1681 accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg 1741 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1801 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1861 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc 1921 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1981 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 2041 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 2101 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 2161 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaattaacaa 2221 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 2281 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 2341 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 2401 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 2461 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 2521 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc 2581 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg 2641 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat 2701 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc 2761 cagtttggtt aattaactgc agccccgata aaataaaaga ttttatttag tctccagaaa 2821 aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagctgcagt aacgccattt 2881 tgcaaggcat ggaaaaatac caaaccaaga atagagaagt tcagatcaag ggcgggtaca 2941 tgaaaatagc taacgttggg ccaaacagga tatctgcggt gagcagtttc ggccccggcc 3001 cggggccaag aacagatggt caccgcagtt tcggccccgg cccgaggcca agaacagatg 3061 gtccccagat atggcccaac cctcagcagt ttcttaagac ccatcagatg tttccaggct 3121 cccccaagga cctgaaatga ccctgcgcct tatttgaatt aaccaatcag cctgcttctc 3181 gcttctgttc gcgcgcttct gcttcccgag ctctataaaa gagctcacaa cccctcactc 3241 ggcgcgccag tcctccgaca gactgagtcg cccgggggtc tagaagccgc caccatgtct 3301 gggagcagct ggcttctcct ctctctcgcg gcactcactg ccgctcaatc tacggaggat 3361 cttgtcaaaa cgttcctcga aaaatttaac tatgaagccg aggaactttc ttaccagtca 3421 tccttggcta gttggaatta caacattaac atcacggatg agaatgtgca gaagatgaat 3481 aatgcagggg ccaagtggtc tgctttctac gaggagcaat ctaaactggc gaaaacctac 3541 ccccttgagg aaatccagga ctcaactgtt aaaaggcaat tgagagcact gcagcattca 3601 ggaagtagtg tgctgtcagc cgacaaaaat cagaggctga acacaatact gaacagcatg 3661 tcaacggtat actcaaccgg taaggcttgt aatccaagca atccgcagga atgcctcctc 3721 cttgagccgg gtctcgacga catcatggag aacagcaagg attacaatga acgcctctgg 3781 gcgtgggaag gctggcggag cgaagtcggc aagcagttgc gccccctcta cgaggaatac 3841 gtagcactca agaacgaaat ggcccgggct aacaactatg aggattacgg agattactgg 3901 aggggggatt atgaggagga gtgggaaaat gggtataatt actccaggaa ccaacttata 3961 gatgacgtgg aacttacgtt cactcagata atgcctcttt atcaacactt gcatgcgtat 4021 gttagaacaa aattgatgga cacatatcct agttatatat ctccgacagg atgtcttcct 4081 gcccatctcc ttggggacat gtgggggcga ttttggacta atctgtatcc actgacagtt 4141 cctttcgggc aaaagcccaa cattgacgtt acgaatgcga tggttaatca atcttgggac 4201 gcccgcaaaa tatttaagga ggccgagaag tttttcgtta gcgtaggcct gccaaatatg 4261 acccaagagt tctggggtaa tagcatgctc accgagccga gtgacagtag gaaagtagta 4321 tgccacccca cggcgtggga ccttgggaaa ggtgattttc gaatcaagat gtgtacgaag 4381 gtcacgatgg acgattttct cactgcacac catgagatgg gccacataca atatgacatg 4441 gcatatgctg cacagccctt tcttttgaga aatggagcga atgagggctt ccacgaggct 4501 gtgggagaga ttatgtcact gtctgcagct actcccaatc acctcaagaa tattgggttg 4561 cttcctccgt ctttcttcga ggattcagag actgaaatta atttcctgct taaacaagcg 4621 ctgacgatcg tcggaactct tccttttacg tacatgctcg agaagtggcg gtggatggtc 4681 ttcaagggtg agatacccaa agatcagtgg atgaaaactt ggtgggagat gaaaaggaac 4741 attgtgggag tagtagaacc tgtcccacat gatgagacat actgtgatcc cgcgtccctc 4801 ttccacgtgg ctaacgatta ctcttttatt cggtattaca cccgcaccat ataccagttc 4861 caattccagg aagctctctg tcagatcgcg aaacacgagg gtcctctcca taaatgtgac 4921 atctcaaaca gcagtgaggc gggccaaaag cttcttgaaa tgttgaaact ggggaagtca 4981 aagccctgga cgtacgctct cgagatagtc gtcggcgcaa agaacatgga tgtacgccct 5041 ctcctcaatt atttcgagcc actcttcaca tggctcaaag aacaaaaccg gaatagtttt 5101 gttgggtgga acaccgattg gagcccgtat gccgaccaat caattaaagt tcggatatcc 5161 cttaagagcg ctctgggcga gaaagcttat gagtggaaca acaatgaaat gtaccttttc 5221 cgctcttcca ttgcctacgc aatgcggcag tatttctcag aagtaaaaaa ccaaaccata 5281 ccattcgttg aggacaatgt gtgggtgtct gatctcaaac cacggatctc atttaacttt 5341 tctgtaacat ctcccggaaa cgtttctgat attatccccc gaactgaggt agaggaggcc 5401 atcagaatgt atagatctcg gatcaatgat gtgtttcgcc tcgacgataa ctcccttgag 5461 tttctcggca tccaaccgac cccgggtccc ccttacgaac ccccggttac gatttggctt 5521 atagtgtttg gtgtagttat gggtgtcgtg gtggttggaa tagttcttct gatattcagc 5581 ggcatacgga ataggcgcaa gaatgaccaa gcacgcggtg aggagaaccc atacgcctcc 5641 gttgacctct caaaggggga aaataacccc ggattccaga gcggggacga tgtacaaaca 5701 tcttttggag cagctgagca gaagctgatc agcgaggagg acctgtagga attctcgacc 5761 ttgactggcg gcgcgacctt gaggcctgcg ttcgcctcag ttgccccctc tgtgcaatgg 5821 ggagacgcgc ctcatcgctt gacaacggcc gaagagccgc cgcgcttccg tctcccgcgt 5881 gcgcgcgcca tgctgcccac ccccgttccg cactgaccct cccccgtgcc ccgcgtcccg 5941 tactgccgcc ccgccccgag tcccatgccg cagccaccgc gacggagccc gcaggcggga 6001 acctgcctcc gcgcgttagc gcgcacgcgc gcctcatgtg tcgtccccat cagcgccggc 6061 ttccgtctat aggccagatg cactgtcact ctggcgaagt cgcagacccg attggccggg 6121 acggaggcgc gagaccgggt tgcgggcggg gccgaacgtg gtataaaagg ggcgggaggc 6181 caggctcgtg ccgttttgca gacgccaccg ccgaggaaaa ccgtgtacta ttagccacgc 6241 gtgccaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc 6301 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg 6361 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg 6421 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca 6481 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt 6541 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc 6601 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg 6661 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg 6721 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg 6781 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg 6841 gtgcctgagc gatcgctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta 6901 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 6961 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 7021 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 7081 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 7141 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 7201 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt 7261 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 7321 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 7381 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 7441 ccccgcttaa tcgcgtcgag acctagaaaa acatggagca atcacaagta gcaatacagc 7501 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc 7561 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 7621 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat 7681 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc 7741 agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 7801 gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag 7861 cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct 7921 agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga 7981 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 8041 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 8101 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttcatgtg 8161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 8221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 8281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 8341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 8401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 8461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 8521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 8581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 8641 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 8701 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 8761 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 8821 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 8881 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 8941 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 9001 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 9061 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 9121 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 9181 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 9241 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 9301 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 9361 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 9421 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 9481 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 9541 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 9601 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 9661 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 9721 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 9781 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 9841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 9901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 9961 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc 10021 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct 10081 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg 10141 aag //