LOCUS Exported 6422 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of human cathepsin B. ACCESSION . VERSION . KEYWORDS hCathepsin B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6422) AUTHORS Huang IC, Bosch BJ, Li F, Li W, Lee KH, Ghiran S, Vasilieva N, Dermody TS, Harrison SC, Dormitzer PR, Farzan M, Rottier PJ, Choe H TITLE SARS coronavirus, but not human coronavirus NL63, utilizes cathepsin L to infect ACE2-expressing cells. JOURNAL J Biol Chem. 2005 Dec 8. ():. PUBMED 16339146 REFERENCE 2 (bases 1 to 6422) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6422 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2015..2032) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2021..2245 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2291..2719 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2378..2397) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2588..2609 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2728..2748) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2733..3062 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2913..3048 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2975..2994 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3183..3202) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3791..3810 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4095..4216 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4132..4151) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4186..4205 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4265..4281) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4265..4281) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4278..4300) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4289..4305 /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(4313..4343) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4358..4379 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4496..4513) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4667..5255) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4747..4766) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5426..6286) /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 6049..6068 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6287..6391) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga atgtggcagc tctgggcctc cctctgctgc ctgctggtgt 961 tggccaatgc ccggagcagg ccctctttcc atcccttaag cgatgagctg gtcaactatg 1021 tcaacaaacg gaataccacg tggcaggccg ggcacaactt ctacaacgtg gacatgagct 1081 acttgaagag gctatgtggt accttcctgg gtgggcccaa gccaccccag agagttatgt 1141 ttaccgagga cctgaagctg cctgcaagct tcgatgcacg ggaacaatgg ccacagtgtc 1201 ccaccatcaa agagatcaga gaccagggct cctgtggcag ctgctgggcc ttcggggctg 1261 tggaagccat ctctgaccgg atctgcatcc acaccaatgc gcacgtcagc gtggaggtgt 1321 cggcggagga cctgctcacc tgctgtggca gcatgtgtgg ggacggctgt aatggtggct 1381 atcctgctga agcttggaac ttctggacaa gaaaaggcct ggtttctggt ggcctctatg 1441 aatcccatgt agggtgcaga ccgtactcca tccctccctg tgagcaccac gtcaacggct 1501 cccggccccc atgcacgggg gagggagata cccccaagtg tagcaagatc tgtgagcctg 1561 gctacagccc gacctacaaa caggacaagc actacggata caattcctac agcgtctcca 1621 atagcgagaa ggacatcatg gccgagatct acaaaaacgg ccccgtggag ggagctttct 1681 ctgtgtattc ggacttcctg ctctacaagt caggagtgta ccaacacgtc accggagaga 1741 tgatgggtgg ccatgccatc cgcatcctgg gctggggagt ggagaatggc acaccctact 1801 ggctggttgc caactcctgg aacactgact ggggtgacaa tggcttcttt aaaatactca 1861 gaggacagga tcactgtgga atcgaatcag aagtggtggc tggaattcca cgcaccgatc 1921 agtactggga aaagatcggt accgagacca gccaggtggc ccccgcctaa ggatccgagc 1981 tcggtaccaa gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag 2041 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2101 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2161 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2221 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2281 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2341 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2401 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2461 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2521 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2581 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2641 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 2701 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2761 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2821 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2881 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2941 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3001 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3061 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3121 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3181 ggctattcgg ctatgactgg gcaacagaca atcggctgct ctgatgccgc cgtgttccgg 3241 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat 3301 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca 3361 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg 3421 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat 3481 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa 3541 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg 3601 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg 3661 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg 3721 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat 3781 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac 3841 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc 3901 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc 3961 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg 4021 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt 4081 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 4141 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 4201 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat 4261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 4321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 4381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 4441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 4501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 4561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 5101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 5221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 5341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 5401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 5521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 5641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 5701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 5761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 5821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 5881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 6001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 6061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 6121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 6181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 6241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 6301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 6361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 6421 tc //