LOCUS       sequence-244739-        7556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse PCSK2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7556)
  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 7556)
  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-mPCSK2
FEATURES             Location/Qualifiers
     source          1..7556
                     /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             3054..3095
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3105..3122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3145..3162)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3151..3375
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3421..3849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3508..3527)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3718..3739
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3858..3878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3863..4192
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4043..4178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4105..4124
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4259..5053
                     /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(4313..4332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4923..4942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5229..5350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5266..5285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5320..5339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5399..5415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5399..5415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5412..5434)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5423..5439
                     /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(5447..5453,5454..5471,5472..5477))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5492..5513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5630..5647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5801..6389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5881..5900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6560..7351,7352..7420))
                     /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     7183..7202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7421..7525)
                     /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 atcttgcatc ttccctcttc gtcccctgct ccaccaccct gcgcgcctca
     1021 cagccccact tttcactccc aaagaaggat ggagggcggt tgtggatccc agtggaaggc
     1081 ggccgggttc ctcttctgtg tgatggtttt tgcgtctgcc gagagacccg tcttcacgaa
     1141 tcattttctt gtggagttgc ataaagacgg agaggaagag gctcgccaag ttgcagcaga
     1201 acacggcttt ggagtccgaa agctcccctt tgcagaaggc ctgtatcact tttatcacaa
     1261 tgggcttgca aaggccaaaa gaagacgcag cctacaccat aagcggcagc tagagagaga
     1321 ccccaggata aagatggcgc tgcaacaaga aggatttgac cgtaaaaaga gagggtacag
     1381 ggacatcaat gagattgaca tcaacatgaa tgatcctctc tttacaaagc aatggtacct
     1441 gttcaacact gggcaagccg atggaactcc tgggctagac ttgaacgtgg ccgaagcctg
     1501 ggagctggga tacacaggaa aaggagtgac cattggaatt atggatgatg gaattgacta
     1561 tctccaccca gacctggcct acaactacaa cgctgatgca agttatgact tcagcagcaa
     1621 tgacccctac ccataccctc gatacacaga tgactggttc aacagccatg gaactaggtg
     1681 tgcaggagaa gtttctgctg cagccagcaa caatatctgt ggagtcggcg tagcatacaa
     1741 ctccaaggtg gcagggatcc ggatgctgga ccagcccttt atgacagaca tcatcgaagc
     1801 ctcctccatc agccacatgc ctcaactgat cgacatctac agtgcaagct ggggccccac
     1861 agacaatggg aagacggttg atgggccccg agagctcaca ctccaggcca tggctgatgg
     1921 cgtgaacaag ggccgtgggg gcaaaggcag catctatgtg tgggcctctg gggacggtgg
     1981 cagctacgat gactgcaact gtgacggcta tgcttcaagc atgtggacca tctccatcaa
     2041 ctcagccatc aatgatggca ggactgcctt gtatgatgag agttgctctt ccaccttagc
     2101 atccaccttc agcaatggga ggaagaggaa tcctgaggct ggtgtggcta ccacagactt
     2161 gtatggcaac tgtactctga gacactctgg gacatctgca gctgctccgg aggcagctgg
     2221 cgtgtttgca ttagctttgg aggctaacct ggatctgacc tggagagaca tgcaacatct
     2281 gactgtgctc acctccaagc ggaaccagct tcatgatgag gttcatcagt ggcgacggaa
     2341 tggggttggc ctggaattta atcacctctt tggctacgga gtccttgatg caggtgccat
     2401 ggtgaaaatg gctaaagact ggaaaactgt tccggagaga ttccattgtg tgggaggctc
     2461 tgtgcagaac cctgaaaaaa taccacccac cggcaagctg gtactgaccc tcaaaacaaa
     2521 tgcatgtgag gggaaggaaa acttcgtccg ctacctggag cacgtccaag ctgtcatcac
     2581 agtcaacgcg accaggagag gagacctgaa catcaacatg acctccccaa tgggcaccaa
     2641 gtccattttg ctgagccggc gtcccagaga cgacgactcc aaggtgggct ttgacaagtg
     2701 gcctttcatg accacccaca cctgggggga ggatgcccga gggacctgga ccctggagct
     2761 ggggtttgtg ggcagtgcac cacagaaggg gttgctgaag gaatggaccc tgatgcttca
     2821 cggcacacag agcgccccat acatcgatca ggtggtgagg gattaccagt ctaagctggc
     2881 catgtccaag aagcaggagc tggaggaaga gctggatgag gctgtggaga gaagcctgca
     2941 aagtatcctg agaaagaact agggccacgc ttccgccttc acctcccctt cctccccgtc
     3001 tctgcctctc cttgctccac agttcctcga gtctagaggg cccgcggttc gaaggtaagc
     3061 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     3121 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3181 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3241 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3301 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3361 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3421 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3481 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3541 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3601 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3661 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3721 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3781 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3841 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3901 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3961 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4021 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4081 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4141 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4201 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4261 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4321 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4381 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4441 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4501 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4561 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4621 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4681 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4741 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4801 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4861 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4921 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4981 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5041 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5101 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5161 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5221 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5281 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5341 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5401 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5461 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5521 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5581 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5641 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5701 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5761 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5821 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5881 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5941 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6001 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6061 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7501 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
