LOCUS Exported 8161 bp ds-DNA circular SYN 16-JUN-2021 DEFINITION Strong human ACE2 abundance from Bxb1 landing pad. ACCESSION . VERSION . KEYWORDS AttB_ACE2_IRES-mCherry-H2A-P2A-PuroR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8161) TITLE Variants of human ACE2 differentially promote SARS-CoV and SARS-CoV-2 spike mediated infection JOURNAL Unpublished REFERENCE 2 (bases 1 to 8161) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8161 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(541..1401) /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 1164..1183 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1402..1506) /gene="bla" /label=AmpR promoter rep_origin complement(1532..1987) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1669..1690) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1881..1900 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2114..2136 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2128..2145 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2129..2145 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2152..2171 /label=T7 /note="T7 promoter, forward primer" promoter 2152..2170 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 4881..5467 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(5048..5065) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5275..5294 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind complement(5613..5631) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 5881..5900 /label=mCherry-F /note="mCherry, forward primer" CDS 6611..6667 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" primer_bind 7161..7181 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 7372..7493 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7409..7428) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7463..7482 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter complement(7502..7520) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7541..7557) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7541..7557) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7554..7576) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7565..7581 /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(7589..7619) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7634..7655 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7772..7789) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7943..370) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8023..8042) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 61 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 121 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 181 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 241 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 301 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 361 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 421 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 481 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 541 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 601 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 661 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 721 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 781 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 841 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 901 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 961 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 1021 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 1081 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 1141 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 1201 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 1261 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 1321 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 1381 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 1441 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 1501 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa 1561 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa 1621 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac 1681 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag 1741 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt 1801 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg 1861 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca 1921 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag 1981 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 2041 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 2101 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac 2161 tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacgatgt aggtcacggc 2221 attccggact cagcgctcga gccggcttgt cgacgacggc ggtctccgtc gtcaggatca 2281 tccgcagatt ataccgcaac tacacgccac catgtccagc tcctcctggc tccttctcag 2341 ccttgttgct gtaactgctg ctcagtccac cattgaggaa caggccaaga catttttgga 2401 caagtttaac cacgaagccg aagacctgtt ctatcaaagt tcacttgctt cttggaatta 2461 taacaccaat attactgaag agaatgtcca aaacatgaat aatgctgggg acaaatggtc 2521 tgccttttta aaggaacagt ccacacttgc ccaaatgtat ccactacaag aaattcagaa 2581 tctcacagtc aagcttcagc tgcaggctct tcagcaaaat gggtcttcag tgctctcaga 2641 agacaagagc aaacggttga acacaattct aaatacaatg agcaccatct acagtactgg 2701 aaaagtttgt aacccagata atccacaaga atgcttatta cttgaaccag gtttgaatga 2761 aataatggca aacagtttag actacaatga gaggctctgg gcttgggaaa gctggagatc 2821 tgaggtcggc aagcagctga ggccattata tgaagagtat gtggtcttga aaaatgagat 2881 ggcaagagca aatcattatg aggactatgg ggattattgg agaggagact atgaagtaaa 2941 tggggtagat ggctatgact acagccgcgg ccagttgatt gaagatgtgg aacatacctt 3001 tgaagagatt aaaccattat atgaacatct tcatgcctat gtgagggcaa agttgatgaa 3061 tgcctatcct tcctatatca gtccaattgg atgcctccct gctcatttgc ttggtgatat 3121 gtggggtaga ttttggacaa atctgtactc tttgacagtt ccctttggac agaaaccaaa 3181 catagatgtt actgatgcaa tggtggacca ggcctgggat gcacagagaa tattcaagga 3241 ggccgagaag ttctttgtat ctgttggtct tcctaatatg actcaaggat tctgggaaaa 3301 ttccatgcta acggacccag gaaatgttca gaaagcagtc tgccatccca cagcttggga 3361 cctggggaag ggcgacttca ggatccttat gtgcacaaag gtgacaatgg acgacttcct 3421 gacagctcat catgagatgg ggcatatcca gtatgatatg gcatatgctg cacaaccttt 3481 tctgctaaga aatggagcta atgaaggatt ccatgaagct gttggggaaa tcatgtcact 3541 ttctgcagcc acacctaagc atttaaaatc cattggtctt ctgtcacccg attttcaaga 3601 agacaatgaa acagaaataa acttcctgct caaacaagca ctcacgattg ttgggactct 3661 gccatttact tacatgttag agaagtggag gtggatggtc tttaaagggg aaattcccaa 3721 agaccagtgg atgaaaaagt ggtgggagat gaagcgagag atagttgggg tggtggaacc 3781 tgtgccccat gatgaaacat actgtgaccc cgcatctctg ttccatgttt ctaatgatta 3841 ctcattcatt cgatattaca caaggaccct ttaccaattc cagtttcaag aagcactttg 3901 tcaagcagct aaacatgaag gccctctgca caaatgtgac atctcaaact ctacagaagc 3961 tggacagaaa ctgttcaata tgctgaggct tggaaaatca gaaccctgga ccctagcatt 4021 ggaaaatgtt gtaggagcaa agaacatgaa tgtaaggcca ctgctcaact actttgagcc 4081 cttatttacc tggctgaaag accagaacaa gaattctttt gtgggatgga gtaccgactg 4141 gagtccatat gcagaccaaa gcatcaaagt gaggataagc ctaaaatcag ctcttggaga 4201 taaagcatat gaatggaacg acaatgaaat gtacctgttc cgatcatctg ttgcatatgc 4261 tatgaggcag tactttttaa aagtaaaaaa tcagatgatt ctttttgggg aggaggatgt 4321 gcgagtggct aatttgaaac caagaatctc ctttaatttc tttgtcactg cacctaaaaa 4381 tgtgtctgat atcattccta gaactgaagt tgaaaaggcc atcaggatgt cccggagccg 4441 tatcaatgat gctttccgtc tgaatgacaa cagcctagag tttctgggga tacagccaac 4501 acttggacct cctaaccagc cccctgtttc catatggctg attgtttttg gagttgtgat 4561 gggagtgata gtggttggca ttgtcatcct gatcttcact gggatcagag atcggaagaa 4621 gaaaaataaa gcaagaagtg gagaaaatcc ttatgcctcc atcgatatta gcaaaggaga 4681 aaataatcca ggattccaaa acactgatga tgttcagacc tcctttggta ccgagacctc 4741 ccaggtggcg cccgcttagt aacttaagaa ttcaccggtc tgacctcctt ctccctctct 4801 tcaggtctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca 4861 ctgtggaatc agagagactg cccctctccc tctccccccc ctaacgttac tggccgaagc 4921 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct 4981 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt 5041 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct 5101 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc 5161 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag 5221 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc 5281 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga 5341 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt 5401 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc 5461 cacaaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg 5521 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg 5581 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc 5641 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt 5701 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg 5761 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct 5821 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg 5881 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga 5941 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta 6001 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta 6061 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca 6121 gtacgaacgc gccgagggcc gccactccac cggcggctac tcagatctcg agctcaagct 6181 tcgaattctg atgtcgggac gcggcaagca gggaggcaaa gctcgcgcca aagccaagac 6241 ccgctcttct cgtgccggtc tccagttccc cgtgggccga gtgcaccgac tgctccgcaa 6301 gggcaactat gctgagcggg tcggggccgg cgcgccggtg tacctggcgg cggtgctgga 6361 gtacctgact gccgagatcc tggagctggc gggcaacgcc gcccgcgaca acaagaagac 6421 ccgcattatc ccgcgccact tgcagctggc catccgcaac gacgaggagc tcaacaagct 6481 gctgggcaaa gtaaccatcg ctcagggtgg tgtcctgccc aacatccagg ctgtgctact 6541 gcccaagaag accgagagtc accacaaggc caaaggcaaa cgggatccac cggatctaga 6601 taacagcgga gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc 6661 tggacctacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc 6721 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc 6781 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 6841 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 6901 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 6961 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 7021 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 7081 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 7141 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 7201 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agtaataggg 7261 atcctgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc 7321 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt 7381 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 7441 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaggagatc 7501 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt 7561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 7621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 7681 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 7741 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cactcggtcg 7801 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 7861 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 7921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 7981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 8041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 8101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 8161 g //