LOCUS       sequence-244744-        7796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse PCSK9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7796)
  AUTHORS   Li JJ, Liu J, Lupino K, Liu X, Zhang L, Pei L
  TITLE     Growth Differentiation Factor 15 Maturation Requires Proteolytic 
            Cleavage by PCSK3, -5, and -6.
  JOURNAL   Mol Cell Biol. 2018 Oct 15;38(21). pii: MCB.00249-18. doi: 
            10.1128/MCB.00249-18. Print 2018 Nov 1.
  PUBMED    30104250
REFERENCE   2  (bases 1 to 7796)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol."; date: "2018-10-15"; volume: "38(21). pii: MCB.00249-18. 
            doi"; pages: " 10.1128/MCB.00249-18. Print 2018 Nov 1"
COMMENT     Sequence Label: pcDNA3.1-mPCSK9
FEATURES             Location/Qualifiers
     source          1..7796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     CDS             1700..1711
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3294..3335
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3345..3362
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3385..3402)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3391..3615
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3661..4089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3748..3767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3958..3979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4098..4118)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4103..4432
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4283..4418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4345..4364
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4499..5293
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4553..4572)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5163..5182
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5469..5590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5506..5525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5560..5579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5639..5655)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5639..5655)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5652..5674)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5663..5679
                     /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(join(5687..5693,5694..5711,5712..5717))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5732..5753
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5870..5887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6041..6629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6121..6140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6800..7591,7592..7660))
                     /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     7423..7442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7661..7765)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttagggcaa
      961 ttctgcagat atctttctga ggctagagga ctgagccagt ccttggctcc ccagagacat
     1021 cacggcccgc agccccggag ccaagtgccc cgagtcccag gcgtccatgt ccttcccgag
     1081 gccgcgcgca cctctcctcg ccccgatggg cacccactgc tctgcgtggc tgcggtggcc
     1141 gctgttgccg ctgttgccgc cgctgctgct gctgttgctg ctactgtgcc ccaccggcgc
     1201 tggtgcccag gacgaggatg gagattatga agagctgatg ctcgccctcc cgtcccagga
     1261 ggatggcctg gctgatgagg ccgcacatgt ggccaccgcc accttccgcc gttgctccaa
     1321 ggaggcctgg aggctgccag gaacctacat tgtggtgctg atggaggaga cccagaggct
     1381 acagattgaa caaactgccc accgcctgca gacccgggct gcccgccggg gctatgtcat
     1441 caaggttcta catatctttt atgacctctt ccctggcttc ttggtgaaga tgagcagtga
     1501 cctgttgggc ctggccctga agttgcccca tgtggagtac attgaggaag actcctttgt
     1561 cttcgcccag agcatcccat ggaacctgga gcgaattatc ccagcatggc accagacaga
     1621 ggaagaccgc tcccctgatg gaagcagcca ggtggaggtg tatctcttag ataccagcat
     1681 ccagggtgcc catcgggaga ttgagggcag ggtcaccatc accgacttca acagcgtgcc
     1741 ggaggaggat gggacacgct tccacagaca ggcgagcaag tgtgacagcc acggcaccca
     1801 cctggcaggt gtggtcagcg gccgggatgc tggtgtggcc aagggcacca gcctgcacag
     1861 cctgcgtgtg ctcaactgtc aagggaaggg cacagtcagc ggcaccctca taggcctgga
     1921 gtttattcgg aagagtcagc taatccagcc ctcggggcca ctcgtggttc tgctgcccct
     1981 ggccggtggg tatagccgca tcctcaacgc tgcctgccgg cacctggcga ggactggggt
     2041 ggtgctggtt gcagcagctg ggaacttccg ggacgacgcc tgcctctact ccccagcttc
     2101 tgctccagag gtcatcacag tcggggccac gaatgcccag gaccagccag ttaccttggg
     2161 gactttgggg actaattttg gacgctgtgt ggatctcttt gcccccggga aggacatcat
     2221 cggagcgtcc agtgactgca gcacatgctt catgtcacag agtgggacct cacaggctgc
     2281 tgcccacgtg gccggcattg tggctcggat gctgagccgg gagcccacac ttaccctggc
     2341 cgagctgcgg cagaggctga tccacttctc taccaaagac gtcatcaaca tggcctggtt
     2401 ccctgaggac cagcaggtgc tgacccccaa cctggtggcc acactgcccc ccagcaccca
     2461 tgagacaggc gggcagctgc tctgtaggac ggtgtggtcg gcacactcgg ggcccactcg
     2521 aacagctaca gctacagccc gctgtgcccc agaagaggag ctgctgagct gctccagctt
     2581 ctccaggagc gggaggcgtc gtggtgattg gattgaggcc ataggaggcc agcaggtctg
     2641 caaggccctc aatgcatttg ggggtgaggg tgtctatgcc gtcgcgagat gctgcctggt
     2701 tccccgtgcc aactgcagca tccacaacac ccctgcagcc agagctggcc tggagaccca
     2761 tgtccactgc caccagaagg accatgttct cacaggctgc agcttccatt gggaagtgga
     2821 agaccttagt gtccggaggc agcctgcgct gaggtccaga cgtcagcctg gccagtgcgt
     2881 tggccaccag gcggccagtg tctatgcttc ctgctgccat gccccagggc tggaatgcaa
     2941 aatcaaggag catgggatct caggtccttc agagcaggtc actgtggcct gcgaagcagg
     3001 atggaccctg actggatgca atgtgctccc tggggcatcc ctcactctgg gagcctacag
     3061 cgtggacaac ctgtgtgtgg caagagtcca tgacactgcc agagcagaca ggaccagtgg
     3121 agaagccaca gtagctgctg ccatctgctg ccggagccgg ccttcagcaa aggcctcctg
     3181 ggttcagtga cagcctcagg cagggatggt gcttgaggct gggtgcagag atatgcctgc
     3241 atggctctct tgtagccaaa ggtggctcga gtctagaggg cccgcggttc gaaggtaagc
     3301 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     3361 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3421 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3481 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3541 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3601 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3661 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3721 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3781 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3841 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3901 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3961 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4021 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4081 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4201 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4261 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4321 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4381 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4441 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4501 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4561 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4621 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4681 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4741 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4801 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4861 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4921 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4981 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5041 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5101 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5161 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5221 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5281 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5341 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5401 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5461 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5521 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5581 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5641 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5701 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5761 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5821 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5881 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5941 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
