LOCUS       sequence-449427-        5629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG-tagged mutated p37 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5629)
  AUTHORS   Wrobel L, Hoffmann JL, Li X, Rubinsztein DC
  TITLE     p37 regulates VCP/p97 shuttling and functions in the nucleus and 
            cytosol.
  JOURNAL   Sci Adv. 2024 May 3;10(18):eadl6082. doi: 10.1126/sciadv.adl6082. 
            Epub 2024 May 3.
  PUBMED    38701207
REFERENCE   2  (bases 1 to 5629)
  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: "Sci Adv."; 
            date: "2024-05-3"; volume: "10(18):eadl6082. doi"; pages: " 
            10.1126/sciadv.adl6082. Epub 2024 May 3"
COMMENT     Sequence Label: pCMV5a-p37SHPmut_FLAG
FEATURES             Location/Qualifiers
     source          1..5629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1005..1024
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1005..1023
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1062..1391)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1076..1211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1376..1396
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1420..2042
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1824..1843
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2050..2073)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3111..3314)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3140..3160)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3315..3694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3855..3872)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3855..3871)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3864..3886)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4012..4467)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4149..4170)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4361..4380
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4494..4598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4599..4667,4668..5459)
                     /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     complement(4817..4836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga catgattacg aatttaatac gactcactat
     1021 agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa gcctaggcct
     1081 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     1141 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     1201 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     1261 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     1321 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     1381 aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg catctactga
     1441 gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc caccggttgg
     1501 gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag ttattttaaa
     1561 atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa tttgggaggc
     1621 caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa tatggtgaaa
     1681 ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc ctggaatccc
     1741 aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga gattgcagtg
     1801 agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca ggccctacag
     1861 gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat aatattatag
     1921 aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag gctggtgggc
     1981 actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca gggatgccac
     2041 ccagtactac ttgtcatcgt cgtccttgta atcggtaccg gatcctttta gttgctggag
     2101 taacacagtg ttaagaatat ctgcttctag cagtgtcagg ctttcatctg ttagctcttt
     2161 attcggaaat gaagtcacaa gaataaagtc aagagccgca aattcaggac gagactgtac
     2221 aataaagttc cggacatcca ggatcctgtg tgtactattg aatctttgta tcaaacgact
     2281 cccatctgct aacctgattt gaatttttgt tgttggcact gaatcatcaa taagaacaac
     2341 tgcattaagt attgatttat cctcctcttc tggagaggaa ggtgtactga ctatttcagg
     2401 tgtaaggctt ccagctgctg cccctgctcc actagcagcc ttgaacctca atctaggttt
     2461 tatgtattct tgatcctgat gatcctccat atccaaattc acttggccac catgaacaag
     2521 gcgctgaagc tccaggggaa tctctcctct cttaacagac tccagaaatt gagcatttgt
     2581 tggttcattg taaggtctca attctccatc atctaaactg aaaccattgc tccacagttt
     2641 aagcaaaatc tgaacatctt gcagctgatt ttctccatag atatattcag accgcttaca
     2701 aaaagaacta cccaatctgt atcctccacc tgtaaatgac ttagatttat catcacctga
     2761 agctcttgtg gcttcattca gagggacagc cccatgttcc cttgcctctt tgaaaagttc
     2821 attcacaatt ttcccagttg aaggtcgaac tatatttaat ccactgtatt catgttcact
     2881 tgagtaaaac cgttgaggtg gtgtccgtgg gctcttgaag actgtggctt taggtctatt
     2941 agacttggaa gatttgcact tcacctcatc ttcatacaat tctgccaagg ccaactgcaa
     3001 atcccgcgcg ctcggaggcc gcggcccgga agacctcctc tcctgctcgc cgggctcagg
     3061 gcctccgccc tccgccatcg caagcttgaa ttctgacggt tcactaaacg agctctgctt
     3121 atatagacct cccaccgtac acgcctaccg cccatttgcg tcaacggggc ggggttatta
     3181 cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg
     3241 gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt ggtgtactgc
     3301 caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga
     3361 aagtcccgta aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg
     3421 tcaatagggg gcggacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac
     3481 cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca
     3541 tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt
     3601 accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt
     3661 gattactatt aataactagt caataatcaa tgtcaacatg gcggtcatat tggacatgag
     3721 ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca atagccaata
     3781 ttgatttatg ctatataacc aatgactaat atggctaatt gccaatattg attcaatgta
     3841 tagatcagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3901 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3961 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa
     4021 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4081 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4141 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4201 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4261 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4321 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4381 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4441 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4501 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4561 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4621 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4681 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4741 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4801 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4861 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4921 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4981 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5041 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5101 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5161 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5221 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5281 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5341 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5401 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5461 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5521 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5581 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
