LOCUS       sequence-244738-        7870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse PCSK1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7870)
  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 7870)
  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-mPCSK1
FEATURES             Location/Qualifiers
     source          1..7870
                     /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             3368..3409
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3419..3436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3459..3476)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3465..3689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3735..4163
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3822..3841)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4032..4053
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4172..4192)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4177..4506
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4357..4492
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4419..4438
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4573..5367
                     /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(4627..4646)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5237..5256
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5543..5664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5580..5599)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5634..5653
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5713..5729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5713..5729)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5726..5748)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5737..5753
                     /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(5761..5767,5768..5785,5786..5791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5806..5827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5944..5961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6115..6703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6195..6214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6874..7665,7666..7734))
                     /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     7497..7516
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7735..7839)
                     /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 atccccttcc tacttgtgtg agaacaaggt tttgagccat ggagcaaaga
     1021 ggttggactc tgcagtgtac tgctttcgcc ttcttttgcg tttggtgtgc actaaacagt
     1081 gtaaaagcaa agaggcagtt tgttaatgaa tgggcggcgg agatccccgg agggcaagaa
     1141 gctgcctctg ccatcgccga agaactgggg tatgaccttt tgggtcagat tggatcactt
     1201 gaaaatcact atttattcaa acacaaaagc catcctcgga ggtcccgaag aagcgctctt
     1261 catatcacta agaggttatc tgatgatgat cgtgtgacgt gggctgaaca acagtatgaa
     1321 aaagagagaa gtaaacgttc agttcaaaaa gactcagcat tggatctctt caatgatcca
     1381 atgtggaatc agcagtggta cttgcaagat accagaatga ctgcagctct gcccaagctg
     1441 gaccttcatg taatacctgt ttgggaaaag ggtattactg gcaaaggagt tgttattact
     1501 gtactggatg atggcttgga gtggaatcac acagacattt atgccaatta tgatccagag
     1561 gctagctatg attttaacga taatgatcat gatccatttc cccgatatga tctcacaaat
     1621 gaaaacaaac atggaacaag atgtgcaggt gaaattgcca tgcaagcaaa taatcacaag
     1681 tgtggggttg gagttgcata taattccaaa gttggaggca taagaatgct ggatggcatt
     1741 gtgactgatg ccattgaggc tagttcaatt ggattcaacc ctggccatgt ggatatttac
     1801 agtgcaagct ggggccctaa tgatgatgga aaaactgtgg aggggcctgg cagactagcc
     1861 cagaaggcat ttgaatatgg tgtcaaacag gggagacaag ggaaaggctc catctttgtc
     1921 tgggcttcag ggaatggggg tcgtcaggga gataactgtg actgtgatgg ctacacagac
     1981 agcatttaca ccatctctat cagcagtgcc tcccagcaag gcctgtcacc ttggtatgca
     2041 gagaagtgtt cttccacatt ggctacctcc tacagcagtg gtgattacac agaccagcga
     2101 ataacaagcg ctgacctgca caatgactgc acagagaccc acacaggcac ctcggcttca
     2161 gcacccctgg ctgctggtat ctttgctctg gccttggagg caaacccaaa tcttacctgg
     2221 agagatatgc agcatctggt tgtctggacc tctgagtacg acccattggc cagtaaccca
     2281 ggttggaaaa agaatggggc aggcttgatg gtgaacagcc gatttggatt tggcttgcta
     2341 aatgccaaag ctctggtgga tttggctgat cctaggacct ggagaaatgt gcctgagaag
     2401 aaagaatgtg ttgtaaaaga caataacttt gagcctagag ccctgaaagc taatggagaa
     2461 gtaattgttg aaatcccaac aagagcttgt gaaggacaag aaaatgctat caagtctctg
     2521 gaacatgtgc aatttgaagc aacaattgaa tattctcgta gaggagacct tcatgtcaca
     2581 ctcacttctg ctgttggaac cagcactgta ctgttggctg aaagggaaag agatacatcc
     2641 cccaatggct ttaagaattg ggacttcatg tctgttcata catggggaga gaatcctgta
     2701 ggcacctgga cattgaaaat tacagacatg tctggaagaa tgcaaaatga aggaaggatt
     2761 gtgaactgga agttgatttt gcatgggaca tcttctcaac cagagcacat gaagcagcct
     2821 cgtgtgtaca catcctacaa tacagtccag aatgacagga gaggagtgga aaagatggtg
     2881 aatgttgtgg agaagcggcc cacacaaaag agcctgaatg gcaatctcct ggtacccaaa
     2941 aactccagca gcagcaatgt ggagggtaga agggatgagc aggtacaagg aactccttca
     3001 aaggccatgc tgcgactcct acaaagtgct tttagcaaga atgcactttc aaaacaatca
     3061 ccaaagaagt ctccaagtgc aaagctcagc atcccttatg aaagtttcta tgaagccttg
     3121 gaaaagctca acaagccctc caagcttgaa ggctctgaag acagtctgta cagtgactat
     3181 gttgatgtat tctataacac aaaaccttat aagcatagag atgacaggct gctgcaagct
     3241 ctcatggaca tcctaaatga ggagaattaa aataagtgtg ttgcctgagt tgggaatctt
     3301 catgcaccct ccttttcctt aaatgttcct gggtctggac tcgagtctag agggcccgcg
     3361 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     3421 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3481 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3541 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3601 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3661 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3721 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3781 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3841 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3901 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3961 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4021 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4081 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4141 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4201 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4261 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4321 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4381 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4441 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4501 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4561 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4621 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4681 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4741 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4801 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4861 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4921 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4981 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5041 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5101 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5161 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5221 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5281 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5341 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5401 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5461 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5521 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5641 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5701 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5761 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5821 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5881 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6121 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6541 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6721 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6781 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6841 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6901 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6961 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7021 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7081 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7141 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7201 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7261 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7321 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7381 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7441 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7501 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7561 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7621 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7681 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7741 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7801 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7861 acctgacgtc
//
