LOCUS       sequence-340468-        6065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PGRMC2 containing an N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  AUTHORS   Chen YJ, Knupp J, Arunagiri A, Haataja L, Arvan P, Tsai B
  TITLE     PGRMC1 acts as a size-selective cargo receptor to drive ER-phagic 
            clearance of mutant prohormones.
  JOURNAL   Nat Commun. 2021 Oct 13;12(1):5991. doi: 10.1038/s41467-021-26225-8.
  PUBMED    34645803
REFERENCE   2  (bases 1 to 6065)
  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: "Nat 
            Commun."; date: "2021-10-13"; volume: "12(1):5991. doi"; pages: " 
            10.1038/s41467-021-26225-8"
COMMENT     Sequence Label: pCMV-FLAG-PGRMC2
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(60..79)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        251..630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        631..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     785..805
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     879..898
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        879..897
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             917..940
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      945..954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1634..1652)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1635..1652)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1672..1689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1678..1902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1948..2376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2035..2054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2245..2266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2385..2405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2390..2719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2570..2705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2632..2651
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2786..3580
                     /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(2840..2859)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3450..3469
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3754..3875
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3791..3810)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3845..3864
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3924..3940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3924..3940)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3937..3959)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3948..3964
                     /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(3972..3978,3979..3996,3997..4002))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4017..4038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4155..4172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4326..4914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4406..4425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5085..5876,5877..5945))
                     /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     5708..5727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5946..6050)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
       61 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
      121 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      181 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      241 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      301 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      421 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      481 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      541 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      601 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      661 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      721 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      781 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      841 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      901 acccaagctt accatggact acaaggacga cgatgacaag ggatccatgg cggctggtga
      961 tggggacgtg aagctaggca ccctggggag tggcagcgag agcagcaacg acggcggcag
     1021 cgagagtcca ggcgacgcgg gagcggcagc ggaaggggga ggctgggcgg cggcggcgtt
     1081 ggcgcttctg acggggggcg gggaaatgct gctgaacgtg gcgctggtgg ctctggtgct
     1141 gctgggggcc taccggctgt gggtgcgctg ggggcggcgg ggtctggggg ccggggccgg
     1201 ggcgggcgag gagagccccg ccacctctct gcctcgcatg aagaagcggg acttcagctt
     1261 ggagcagctg cgccagtacg acggctcccg caacccgcgc atcctgctcg cggtcaatgg
     1321 gaaagtcttc gacgtgacca aaggcagcaa gttctacggc ccggcgggtc catatggaat
     1381 atttgctggt agggatgcct ccagaggact ggccacattt tgcctagata aagatgcact
     1441 tagagatgaa tatgatgatc tctcagattt gaatgcagta caaatggaga gtgttcgaga
     1501 atgggaaatg cagtttaaag aaaaatatga ttatgtaggc agactcctaa aaccaggaga
     1561 agaaccatca gaatatacag atgaagaaga taccaaggat cacaataaac aggattgatc
     1621 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     1681 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1741 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1801 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1861 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1921 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     1981 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2041 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2101 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2161 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2221 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2281 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2341 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2401 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2461 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2521 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2581 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2641 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     2701 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     2761 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     2821 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     2881 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     2941 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3001 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3061 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3121 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3181 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3241 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3301 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3361 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3421 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3481 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3541 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     3601 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     3661 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     3721 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     3781 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3841 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     3901 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3961 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4021 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4081 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4141 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4201 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4261 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4321 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4381 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4441 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4501 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4561 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4621 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4681 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4741 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4801 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4861 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4921 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4981 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5041 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5101 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5161 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5221 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5281 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5341 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5401 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5461 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5521 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5581 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5641 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5701 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5761 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5821 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5881 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5941 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6001 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6061 cgaaa
//
