LOCUS       sequence-244737-        7480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse PCSK4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7480)
  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 7480)
  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-mPCSK4
FEATURES             Location/Qualifiers
     source          1..7480
                     /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             2978..3019
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3029..3046
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3069..3086)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3075..3299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3345..3773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3432..3451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3642..3663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3782..3802)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3787..4116
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3967..4102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4183..4977
                     /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(4237..4256)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4847..4866
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5153..5274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5190..5209)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5244..5263
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5323..5339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5323..5339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5336..5358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5347..5363
                     /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(5371..5377,5378..5395,5396..5401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5416..5437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5554..5571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5725..6313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5805..5824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6484..7275,7276..7344))
                     /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     7107..7126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7345..7449)
                     /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 gatcctatgc ggccctccca gacagagctg tggctgggtc
      961 tgactttgac tttggccctc ctggctgtga ggtgggcctc agcccaggcc cccatctatg
     1021 tcagcagctg ggcagtgcgg gtgaccaaag gttaccagga agctgagcgc ctggcccgta
     1081 aatttggctt cgtcaacctg ggacagatct tccctgacga ccagtatttc catctgcggc
     1141 accggggtgt ggcccagcag tccctgactc cacactgggg ccatcgtcta cgactgaaga
     1201 aagatcccaa ggtgcgctgg tttgagcagc agactttgag gcgccgggtg aaacgctccc
     1261 tggtggtgcc cacagacccc tggttttcca agcagtggta catgaacaag gagatacaac
     1321 aagatctcaa catcctaaag gcttggaacc agggactgac cggccgggga gtggtgattt
     1381 ccatcttgga cgatggcatt gagaaagacc acccggacct ctgggctaat tatgaccctc
     1441 tggccagcta tgacttcaac gactatgacc cagaccccca gcctcgctac acacccaacg
     1501 atgagaaccg acatggaacg cgctgtgctg gggaggtgtc tgccacagct aacaacggtt
     1561 tctgtggtgc cggtgtggcc ttcaatgcta gaattggagg cgtgcgcatg ttggatggag
     1621 ccatcactga catcgtggag gctcagtccc tcagcctgca gccgcaacac atacacatct
     1681 atagcgccag ctggggcccg gaggatgatg gtcgcacggt ggacggacct ggcctcctca
     1741 cacaggaggc cttcaggcgt ggtgtaacca agggccgcca agggctgggt acactgttca
     1801 tctgggcctc gggaaacggt ggcctccatt acgacaactg caattgtgac ggctacacca
     1861 acagcatcca cacactgtca gtgggcagca ccacgcggca gggtcgagtg ccctggtaca
     1921 gcgaggcctg tgcctccacg tttaccacca ccttcagcag cggtgtggtc accgacccac
     1981 agatcgtcac cacggacctg caccaccagt gcaccgacaa gcacacgggc acctcggcat
     2041 ctgccccgct ggctgctggc atgatcgccc tggccctgga ggccaacccg ctcctgacct
     2101 ggagggacct acagcacctg gtggtccgcg cgtccaggcc ggcgcagctg caggcggagg
     2161 actggaggat caacggcgtg gggcgccaag tgagccacca ttatggctat gggctgctgg
     2221 acgcggggtt gctggtggac ctggctcgcg tgtggctgcc cactaagcct cagaagaaat
     2281 gtgccattcg ggtggtgcac acccccaccc ccatcctgcc tcggatgctg gtgccgaaga
     2341 acgtgaccgc gtgctcagat ggctcgcgcc gccgcctcat ccgctcgctc gagcacgtgc
     2401 aggtccagct ttcgctctcc tacagccgcc gcggggacct ggagatcttc ctcaccagcc
     2461 ccatgggcac gcgctccacg ctcgtggcca tcagaccgtt ggatatcagc ggccaaggct
     2521 acaacaactg gatcttcatg tccactcact actgggatga ggacccgcag ggcctgtgga
     2581 ccctgggcct ggagaataag ggctactatt ttaacacagg aactctgtac tactacaccc
     2641 tgctgctgta tgggacggcc gaggacatga cagcacggcc ccaggccccc caggtgacca
     2701 gccgcgcgcg cgcatgtgtg cagagggaca cagaggggct gtgccaggaa agtcacagtc
     2761 ccctctccat cctggcagga ctctgcctca tctccagcca gcagtggtgg tggctctaca
     2821 gccacccaca gcagccagtg accgaaggac aggccagctg tcaccctcct gttacacctg
     2881 ctgcagcagc ctgagcagcg actacactgc ctgtcccctg tcccttgtgt tgggagagct
     2941 ccgcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct
     3001 cctcggtctc gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg
     3061 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3121 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3181 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3241 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3301 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4141 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4201 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4261 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4321 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4381 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4441 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4501 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4561 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4621 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4681 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4741 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4801 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4861 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4921 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4981 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5041 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5101 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5161 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5221 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5281 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5341 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5461 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
