LOCUS       Exported                6395 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of human cathepsin L.
ACCESSION   .
VERSION     .
KEYWORDS    hCathepsin L
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6395)
  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 6395)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6395
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        80..459
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        460..663
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     614..634
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     708..727
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        708..726
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1833..1850)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1839..2063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2109..2537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2196..2215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2406..2427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2546..2566)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2551..2880
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2731..2866
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2793..2812
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3001..3020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3609..3628
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3913..4034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3950..3969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4004..4023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4083..4099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4083..4099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4096..4118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4107..4123
                     /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(4131..4161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4176..4197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4314..4331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4485..5073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4565..4584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5244..6104)
                     /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     5867..5886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6105..6209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6284..6303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
       61 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
      121 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      181 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      241 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      301 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      361 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      421 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      481 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      541 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      601 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      661 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
      721 tatagggaga cccaagctgg ctagcgttta aacgggccct ctagaatgaa tcctacactc
      781 atccttgctg ccttttgcct gggaattgcc tcagctactc taacatttga tcacagttta
      841 gaggcacagt ggaccaagtg gaaggcgatg cacaacagat tatacggcat gaatgaagaa
      901 ggatggagga gagcagtgtg ggagaagaac atgaagatga ttgaactgca caatcaggaa
      961 tacagggaag ggaaacacag cttcacaatg gccatgaacg cctttggaga catgaccagt
     1021 gaagaattca ggcaggtgat gaatggcttt caaaaccgta agcccaggaa ggggaaagtg
     1081 ttccaggaac ctctgtttta tgaggccccc agatctgtgg attggagaga gaaaggctac
     1141 gtgactcctg tgaagaatca gggtcagtgt ggttcttgtt gggcttttag tgctactggt
     1201 gctcttgaag gacagatgtt ccggaaaact gggaggctta tctcactgag tgagcagaat
     1261 ctggtagact gctctgggcc tcaaggcaat gaaggctgca atggtggcct aatggattat
     1321 gctttccagt atgttcagga taatggaggc ctggactctg aggaatccta tccatatgag
     1381 gcaacagaag aatcctgtaa gtacaatccc aagtattctg ttgctaatga caccggcttt
     1441 gtggacatcc ctaagcagga gaaggccctg atgaaggcag ttgcaactgt ggggcccatt
     1501 tctgttgcta ttgatgcagg tcatgagtcc ttcctgttct ataaagaagg catttatttt
     1561 gagccagact gtagcagtga agacatggat catggtgtgc tggtggttgg ctacggattt
     1621 gaaagcacag aatcagataa caataaatat tggctggtga agaacagctg gggtgaagaa
     1681 tggggcatgg gtggctacgt aaagatggcc aaagaccgga gaaaccattg tggaattgcc
     1741 tcagcagcca gctaccccac tgtgggtacc gagacctccc aggtggcgcc cgcttaagct
     1801 tggatccgcg ttaagtttaa accgctgatc agcctcgact gtgccttcta gttgccagcc
     1861 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1921 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1981 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2041 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2101 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2161 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2221 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2281 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2341 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2401 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2461 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2521 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2701 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     2761 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     2821 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     2881 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     2941 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3001 ctattcggct atgactgggc aacagacaat cggctgctct gatgccgccg tgttccggct
     3061 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3121 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3181 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3241 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3301 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3361 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3421 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3481 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3541 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3601 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3661 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3721 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3781 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3841 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3901 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3961 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4021 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4081 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4141 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4201 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4261 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4321 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4381 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4981 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5041 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5101 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5161 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5221 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5281 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5341 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5401 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5461 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5521 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5581 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5641 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5701 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5761 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5821 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5881 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5941 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6001 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6061 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6121 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6181 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     6241 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     6301 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     6361 cgagcaaaat ttaagctaca acaaggcaag gcttg
//
