LOCUS       sequence-244741-        8784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse PCSK8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8784)
  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 8784)
  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-mPCSK8
FEATURES             Location/Qualifiers
     source          1..8784
                     /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"
     regulatory      4087..4096
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4282..4323
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4333..4350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4373..4390)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4379..4603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4649..5077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4736..4755)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4946..4967
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5086..5106)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5091..5420
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5271..5406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5487..6281
                     /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(5541..5560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6151..6170
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6457..6578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6494..6513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6548..6567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6627..6643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6627..6643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6640..6662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6651..6667
                     /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(6675..6681,6682..6699,6700..6705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6720..6741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6858..6875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7029..7617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7109..7128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7788..8579,8580..8648))
                     /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     8411..8430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8649..8753)
                     /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 atcccagaac agaaacccac ttctgatcaa ggaagagcct cgtgcgattt
     1021 gaatttatgc aattttctga ccatattcac tgaggaccat gaagctcgtc agcacctggc
     1081 ttcttgtgct ggtggttttg ctctgtggga aacggcacct gggcgacagg ctggggacga
     1141 gagctttgga aaaggccccg tgccccagct gctcccacct gactttgaag gtggaattct
     1201 cttcaactgt ggtggagtac gaatatattg tggctttcaa cggatacttc acagccaaag
     1261 ctagaaactc atttatttca agtgctctga aaagcagtga agtggaaaac tggagaataa
     1321 tacctcggaa caacccatcc agtgactacc ctagtgattt tgaggtgatt cagataaaag
     1381 agaagcagaa ggcggggctg ctcacacttg aagatcaccc caacatcaag cgggtgacac
     1441 cccagcggaa agtctttcgt tccctcaagt ttgctgaatc caaccccatc gtgccctgta
     1501 atgaaacccg gtggagccag aagtggcagt catcacgtcc cctgaaaaga gccagtctct
     1561 ccctgggctc tggattctgg catgcaacag gaagacattc aagtcggcga ttgctgagag
     1621 ccattcctcg ccaggtcgcc cagacactgc aggcagatgt gctgtggcag atgggataca
     1681 caggtgctaa tgtcagagtt gctgtttttg atactgggct cagtgagaag catccgcatt
     1741 ttaagaatgt gaaggagaga accaactgga ccaatgagcg gaccctggat gatgggctag
     1801 gccatggcac attcgttgca ggtgtgattg ccagcatgag ggagtgccaa ggatttgctc
     1861 cagatgcaga gctgcacatc ttcagggtct ttaccaacaa tcaggtgtct tacacatctt
     1921 ggtttctgga tgccttcaac tatgccatcc taaagaagat ggacgttctc aaccttagca
     1981 tcggtgggcc cgacttcatg gatcatccgt ttgttgacaa ggtgtgggaa ttaacagcta
     2041 acaatgtaat tatggtttct gctattggca atgatggacc tctctatggc actctgaata
     2101 accctgctga tcagatggat gtgattggag tgggtggcat tgactttgaa gataacatcg
     2161 ctcgcttttc ttccagggga atgactacct gggaattacc aggaggctat ggtcgtgtga
     2221 agcctgacat tgtcacctat ggtgctggag tgcggggttc cggtgtgaaa gggggctgcc
     2281 gtgcactctc agggaccagt gtcgcttccc cagtggtcgc tggggccgtc accttgttag
     2341 taagcacagt acagaagcgg gagctggtga atcctgccag tgtgaagcaa gctttgatag
     2401 cgtcagcccg gagacttcct ggggtcaaca tgttcgagca aggtcatggc aagttggatc
     2461 tgctgcgagc ttatcagatc ctcagcagct ataaaccgca ggcaagcctg agtcctagct
     2521 acatcgacct gactgagtgt ccctacatgt ggccctactg ctcccagcct atctactatg
     2581 gaggaatgcc aacaatcgtt aatgtcacca tcctcaatgg catgggcgtc acaggaagaa
     2641 ttgtggataa gcctgagtgg cgaccctatt taccacagaa tggagacaac attgaagtgg
     2701 ccttctccta ctcctcagtg ttgtggccct ggtcaggtta ccttgccatc tccatttctg
     2761 tgaccaagaa ggcagcttcc tgggaaggca tcgctcaggg ccacatcatg atcacagtgg
     2821 cgtccccagc agagacagag ttacacagtg gtgcggagca cacttccacc gtgaagctgc
     2881 ccatcaaggt gaagatcatt cccacccctc ctcggagcaa gagagtcctc tgggaccagt
     2941 accacaacct ccgctaccca cctggctact tccccaggga caacttgcgg atgaagaatg
     3001 accctttaga ctggaatggc gaccacgtcc acaccaactt cagggacatg taccagcatc
     3061 tgcgcagcat gggctacttc gtggaggtgc tcggcgcccc attcacatgt tttgacgcca
     3121 cacagtatgg cactttgctg ctggtggaca gtgaggaaga gtacttccct gaggagattg
     3181 ctaagctgag gagggatgtg gacaatggcc tttccctcgt catcttcagt gactggtaca
     3241 acacttctgt tatgagaaaa gtgaagtttt atgatgaaaa caccaggcag tggtggatgc
     3301 cagacaccgg aggagcgaac atcccagctc tgaatgagct gctgtctgtg tggaacatgg
     3361 ggttcagtga cggcctatat gaaggggagt ttgtcctggc aaaccatgac atgtactatg
     3421 cgtcggggtg cagcatcgcc aagtttccag aagatggcgt cgtgatcaca cagactttca
     3481 aggaccaagg attggaggtc ttaaaacaag agacagcagt tgtggaaaat gttcccattt
     3541 tggggcttta tcagattcca tctgaaggtg gaggccggat cgtgctgtat ggagactcca
     3601 actgcttgga tgacagtcac agacagaagg actgcttttg gcttctggat gcgctccttc
     3661 agtacacatc ctatggcgtg acccctccca gcctcagcca ttcagggaac cggcagcgcc
     3721 cacctagcgg agccggcttg gcccctcctg aaaggatgga aggaaaccac ctccatcggt
     3781 actccaaagt tcttgaagcc cacttgggag acccgaaacc tcggcccctg ccagcctgtc
     3841 cacatttgtc atgggccaag ccacagcctt tgaatgagac ggcacccagt aatctttgga
     3901 aacatcagaa gctgctctcc attgacctgg acaaagtagt gttacccaac tttcgatcca
     3961 atcgccctca agtgagacct ttgtcccctg gagagagtgg tgcctgggac attcctggag
     4021 ggatcatgcc tggccgctac aaccaggagg tgggacagac catccccgtc ttcgccttcc
     4081 tcggagccat ggtggccctg gccttctttg tggtacagat cagcaaggcc aagagccggc
     4141 cgaagcggag gaggcccagg gcaaagcgtc cacaacttgc acagcaggcc caccctgcaa
     4201 ggaccccatc agtgtgagca tcgcagtagc cagccacaga agctaacaag cctctcgagt
     4261 ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     4321 acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     4381 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4441 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4501 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4561 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4621 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4681 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4741 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4801 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4861 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4921 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4981 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5041 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     5101 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5161 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5221 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5281 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5341 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5401 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     5461 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5521 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5581 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5641 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5701 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5761 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5821 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5881 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5941 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6001 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6061 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6121 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6181 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6241 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     6301 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6361 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6421 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     6481 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6541 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6601 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6661 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6721 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6781 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6841 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6901 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6961 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7021 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7081 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7201 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7441 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7921 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8161 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8761 ccccgaaaag tgccacctga cgtc
//
