LOCUS Exported 3729 bp ds-DNA circular SYN 16-JUN-2021 DEFINITION Strong mCherry fluorescence from Bxb1 landing pad. ACCESSION . VERSION . KEYWORDS AttB_mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3729) TITLE Variants of human ACE2 differentially promote SARS-CoV and SARS-CoV-2 spike mediated infection JOURNAL Unpublished REFERENCE 2 (bases 1 to 3729) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3729 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 113..130 /label=L4440 /note="L4440 vector, forward primer" protein_bind 247..268 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 283..313 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 321..337 /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)." primer_bind 326..348 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 345..361 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 345..361 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 382..400 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 409..530 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(420..439) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 474..493 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(644..1354) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(922..941) /label=mCherry-F /note="mCherry, forward primer" primer_bind 1191..1209 /label=mCherry-R /note="mCherry, reverse primer" regulatory 1351..1360 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(1495..1514) /label=T7 /note="T7 promoter, forward primer" promoter complement(1496..1514) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1521..1538) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1521..1537) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1530..1552) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 1679..2134 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1766..1785) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1976..1997 /label=F1ori-F /note="F1 origin, forward primer" promoter 2160..2264 /gene="bla" /label=AmpR promoter CDS 2265..3125 /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 complement(2483..2502) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 61 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagt gcagcgagtc 121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 241 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 301 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 361 ccatgattac gccaagctcg aaattaaccc tcactaaagg ggatctccta agatacattg 421 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 481 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca 541 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt 601 aaaacctcta caaatgtggt atggctgatt atgatcagga tccctacttg tacagctcgt 661 ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta ctgttccacg atggtgtagt 721 cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt gtaggcgccg ggcagctgca 781 cgggcttctt ggccttgtag gtggtcttga cctcagcgtc gtagtggccg ccgtccttca 841 gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc ctcggggtac atccgctcgg 901 aggaggcctc ccagcccatg gtcttcttct gcattacggg gccgtcggag gggaagttgg 961 tgccgcgcag cttcaccttg tagatgaact cgccgtcctg cagggaggag tcctgggtca 1021 cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga 1081 aggacagctt caagtagtcg gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt 1141 acatgaactg aggggacagg atgtcccagg cgaagggcag ggggccaccc ttggtcacct 1201 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga 1261 actcgtggcc gttcacggag ccctccatgt gcaccttgaa gcgcatgaac tccttgatga 1321 tggccatgtt atcctcctcg cccttgctca ccatggtggc gtgtagttgc ggtataatct 1381 gcggatgatc ctgacgacgg agaccgccgt cgtcgacaag ccggctcgag cgctgagtcc 1441 ggaatgccgt gacctacatc gtcgacctcg agggggggcc cggtacccaa ttcgccctat 1501 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 1561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 1621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac 1681 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 1741 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 1801 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 1861 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 1921 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 1981 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 2041 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 2101 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc 2161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 2221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 2281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 2341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 2401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa 2461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 2521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 2581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 2641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 2701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 2761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 2821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 2881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 2941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 3001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 3061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 3121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt 3181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac 3241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 3301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 3361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 3421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga 3481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 3541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 3601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 3661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 3721 aggcggaca //