LOCUS       sequence-450581-        7596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3.1-p97-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7596)
  AUTHORS   Blythe EE, Gates SN, Deshaies RJ, Martin A
  TITLE     Multisystem Proteinopathy Mutations in VCP/p97 Increase NPLOC4.UFD1L
            Binding and Substrate Processing.
  JOURNAL   Structure. 2019 Dec 3;27(12):1820-1829.e4. doi: 
            10.1016/j.str.2019.09.011. Epub 2019 Oct 14.
  PUBMED    31623962
REFERENCE   2  (bases 1 to 7596)
  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"; doi: 
            "10.1016/j.str.2019.09.011"; journalName: "Structure"; date: 
            "2019-12-3- 3"; volume: "27"; issue: "12"; pages: "1820-1829.e4"
COMMENT     Sequence Label: pAM233
FEATURES             Location/Qualifiers
     source          1..7596
                     /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             3344..3367
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3482..3505
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3604..3621)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3610..3834
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3880..4308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3967..3986)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4177..4198
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4322..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4564..4583
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4699..4746
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4765..5139
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5269..5390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5306..5325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5360..5379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5439..5455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5439..5455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5452..5474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5463..5479
                     /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(5487..5493,5494..5511,5512..5517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5532..5553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5670..5687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5841..6429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5921..5940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6600..7391,7392..7460))
                     /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     7223..7242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7461..7565)
                     /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 gctggctagc
      901 gtttaaactt aagcttgcgc gcgccatggc ttctggagcc gattcaaaag gtgatgacct
      961 atcaacagcc attctcaaac agaagaaccg tcccaatcgg ttaattgttg atgaagccat
     1021 caatgaggac aacagtgtgg tgtccttgtc ccagcccaag atggatgaat tgcagttgtt
     1081 ccgaggtgac acagtgttgc tgaaaggaaa gaagagacga gaagctgttt gcatcgtcct
     1141 ttctgatgat acttgttctg atgagaagat tcggatgaat agagttgttc ggaataacct
     1201 tcgtgtacgc ctaggggatg tcatcagcat ccagccatgc cctgatgtga agtacggcaa
     1261 acgtatccat gtgctgccca ttgatgacac agtggaaggc attactggta atctcttcga
     1321 ggtatacctt aagccgtact tcctggaagc gtatcgaccc atccggaaag gagacatttt
     1381 tcttgtccgt ggtgggatgc gtgctgtgga gttcaaagtg gtggaaacag atcctagccc
     1441 ttattgcatt gttgctccag acacagtgat ccactgcgaa ggggagccta tcaaacgaga
     1501 ggatgaggaa gagtccttga atgaagtagg gtatgatgac attggtggct gcaggaagca
     1561 gctagctcag ataaaggaga tggtggaact gcccctgaga catcctgccc tctttaaggc
     1621 aattggtgtg aagcctccta gaggaatcct gctttacgga cctcctggaa caggaaagac
     1681 cctgattgct cgagctgtag caaatgagac tggagccttc ttcttcttga tcaatggtcc
     1741 tgagatcatg agcaaattgg ctggtgagtc tgagagcaac cttcgtaaag cctttgagga
     1801 ggctgagaag aatgctcctg ccatcatctt cattgatgag ctagatgcca tcgctcccaa
     1861 aagagagaaa actcatggcg aggtggagcg gcgcattgta tcacagttgt tgaccctcat
     1921 ggatggccta aagcagaggg cacatgtgat tgttatggca gcaaccaaca gacccaacag
     1981 cattgaccca gctctacggc gatttggtcg ctttgacagg gaggtagata ttggaattcc
     2041 tgatgctaca ggacgcttag agattcttca gatccatacc aagaacatga agctggcaga
     2101 tgatgtggac ctggaacagg tagccaatga gactcacggg catgtgggtg ctgacttagc
     2161 agccctgtgc tcagaggctg ctctgcaagc catccgcaag aagatggatc tcattgacct
     2221 agaggatgag accattgatg ccgaggtcat gaactctcta gcagttacta tggatgactt
     2281 ccggtgggcc ttgagccaga gtaacccatc agcactgcgg gaaaccgtgg tagaggtgcc
     2341 acaggtaacc tgggaagaca tcgggggcct agaggatgtc aaacgtgagc tacaggagct
     2401 ggtccagtat cctgtggagc acccagacaa attcctgaag tttggcatga caccttccaa
     2461 gggagttctg ttctatggac ctcctggctg tgggaaaact ttgttggcca aagccattgc
     2521 taatgaatgc caggccaact tcatctccat caagggtcct gagctgctca ccatgtggtt
     2581 tggggagtct gaggccaatg tcagagaaat ctttgacaag gcccgccaag ctgccccctg
     2641 tgtgctattc tttgatgagc tggattcgat tgccaaggct cgtggaggta acattggaga
     2701 tggtggtggg gctgctgacc gagtcatcaa ccagatcctg acagaaatgg atggcatgtc
     2761 cactaaaaaa aatgtgttca tcattggcgc taccaaccgg cctgacatca ttgatcctgc
     2821 catcctcaga cctggccgtc ttgatcagct catctacatc ccacttcctg atgagaagtc
     2881 ccgtgttgcc atcctcaagg ctaacctgcg caagtcccca gttgccaagg atgtggactt
     2941 ggagttcctg gctaaaatga ctaatggctt ctctggagct gacctgacag agatttgcca
     3001 gcgtgcttgc aagctggcca tccgtgaatc catcgagagt gagattaggc gagaacgaga
     3061 gaggcagaca aacccatcag ccatggaggt agaagaggat gatccagtgc ctgagatccg
     3121 tcgagatcac tttgaagaag ccatgcgctt tgcgcgccgt tctgtcagtg acaatgacat
     3181 tcggaagtat gagatgtttg cccagaccct tcagcagagt cggggctttg gcagcttcag
     3241 attcccttca gggaaccagg gtggagctgg ccccagtcag ggcagtggag gcggcacagg
     3301 tggcagtgta tacacagaag acaatgatga tgacctgtat ggcgattata aagatgatga
     3361 tgataaataa tcggatccac tagtccagtg tggtggaatt ctgcagatat ccagcacagt
     3421 ggcgggaggc ggcacaggtg gcagtgtata cacagaagac aatgatgatg acctgtatgg
     3481 cgattataaa gatgatgatg ataaataatc ggatccacta gtccagtgtg gtggaattct
     3541 gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat
     3601 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3661 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3721 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3781 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3841 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4081 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4201 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4261 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4681 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     4741 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     4801 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     4861 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     4921 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     4981 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     5041 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     5101 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     5161 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5221 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5281 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5341 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5401 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5461 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5521 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5581 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5641 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5701 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5761 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5821 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5881 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5941 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6001 gcctttctcc cttcgggaag cgtggcgctt tctcgtagct cacgctgtag gtatctcagt
     6061 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6481 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6541 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6601 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6661 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6721 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6781 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6841 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6901 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6961 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7021 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7081 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7141 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7201 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7261 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7321 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7381 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7441 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7501 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7561 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
