LOCUS       sequence-449429-        8760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mClover2-tagged p37 protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8760)
  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 8760)
  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: pEF-1-alpha-p37-mClover2
FEATURES             Location/Qualifiers
     source          1..8760
                     /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"
     LTR             651..831
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     1340..1356
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1341..1357
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1360..1948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1466..1477
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1875..1895
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2042..2752,2753..2755))
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNSIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2084..2105)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2692..2713
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     intron          3783..4721
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3806..3826)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(4722..4933)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    5036..5153
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5451..5495)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5680..5913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6410..6535
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6582..6762
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6780..6978)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6805..6825)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6980..7359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7531..7550
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7625..7729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7730..7798,7799..8590)
                     /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(7948..7967)
                     /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 gctcacatgc tgctagagat
      661 tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg
      721 ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc
      781 tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc cagtacagtc
      841 cggatgcagc tctcgggcca tgtgatgaaa tgctaggcgg ctgtcaaacc tccactctaa
      901 tacttctctc tccgggtcat ccatcccatg caggctcaca gggtgtaaca agcgggtgtt
      961 ctctccttca ttggcttctt ctaccttctc ttgctcaact ggtactagct tgtagcacca
     1021 tccaaaggtc agtggatatc tgatccctgg ccctggtgtg tagttctgcc aatcagggaa
     1081 gtagccttgt gtgtggtaga tccacagatc aaggatatct tgtcttcgtt gggagtgaat
     1141 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     1201 agtcattggt cttaaaggta cctgaggtgt gactggaaaa cccacctcct cctcctcttg
     1261 tgcttctagc caggcacaat cagcattggt agctgctgta ttgctacttg tgattgctcc
     1321 atgtttttct aggtctcgat cgaggtcgac ggtatcgatg cggggaggcg gcccaaaggg
     1381 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     1441 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     1501 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     1561 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1621 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     1681 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1741 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1801 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1861 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1921 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tgtacctagt
     1981 ggaaccggaa cccttaaaca tgtctcgacc tggcttacta gtcggttcaa ctctaggttt
     2041 acttgtacag ctcgtccatg ccatgtgtaa tcccggcggc ggtcacgaac tccagcagga
     2101 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgatgg ctcaggtagt
     2161 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     2221 cgagctgcac gctgccgtcc tcaacgttgt ggcggatctt gaagttagcc ttgatgctgt
     2281 tcttctgctt gtcggccgtg atatagacgt agtggctgtt gaagttgtac tccagcttgt
     2341 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     2401 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta ggtaccgtcg tccttgaaag
     2461 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     2521 tcatgtggtc ggggtagcgg ctgaagcagg ccacgccgta gccgaaggtg gtcacgaggg
     2581 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     2641 tggtggcatc gccctcgccc tcgccgcgga cgctgaactt gtggccgttt acgtcgccgt
     2701 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcactgcgg
     2761 ttggcgaccg gtggatccct tttagttgct ggagtaacac agtgttaaga atatctgctt
     2821 ctagcagtgt caggctttca tctgttagct ctttattcgg aaatgaagtc acaagaataa
     2881 agtcaagagc cgcaaattca ggacgagact gtacaataaa gttccggaca tccaggatcc
     2941 tgtgtgtact attgaatctt tgtatcaaac gactcccatc tgctaacctg atttgaattt
     3001 ttgttgttgg cactgaatca tcaataagaa caactgcatt aagtattgat ttatcctcct
     3061 cttctggaga ggaaggtgta ctgactattt caggtgtaag gcttccaagt ttttgccctt
     3121 ctccactaaa agccttgaac ctcaatctag gttttatgta ttcttgatcc tgatgatcct
     3181 ccatatccaa attcacttgg ccaccatgaa caaggcgctg aagctccagg ggaatctctc
     3241 ctctcttaac agactccaga aattgagcat ttgttggttc attgtaaggt ctcaattctc
     3301 catcatctaa actgaaacca ttgctccaca gtttaagcaa aatctgaaca tcttgcagct
     3361 gattttctcc atagatatat tcagaccgct tacaaaaaga actacccaat ctgtatcctc
     3421 cacctgtaaa tgacttagat ttatcatcac ctgaagctct tgtggcttca ttcagaggga
     3481 cagccccatg ttcccttgcc tctttgaaaa gttcattcac aattttccca gttgaaggtc
     3541 gaactatatt taatccactg tattcatgtt cacttgagta aaaccgttga ggtggtgtcc
     3601 gtgggctctt gaagactgtg gctttaggtc tattagactt ggaagatttg cacttcacct
     3661 catcttcata caattctgcc aaggccaact gcaaatcccg cgcgctcgga ggccgcggcc
     3721 cggaagacct cctctcctgc tcgccgggct cagggcctcc gccctccgcc atctcacgac
     3781 acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat gaaccaagat
     3841 ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc tggcctaact
     3901 tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc cccccaacct
     3961 aaagacgacg tactccaaaa gctcgagaac taatcgaggt gcctggacgg cgcccggtac
     4021 tccgtggagt cacatgaagc gacggctgag gacggaaagg cccttttcct ttgtgtgggt
     4081 gactcacccg cccgctctcc cgagcgccgc gtcctccatt ttgagctccc tgcagcaggg
     4141 ccgggaagcg gccatctttc cgctcacgca actggtgccg accgggccag ccttgccgcc
     4201 cagggcgggg cgatacacgg cggcgcgagg ccaggcacca gagcaggccg gccagcttga
     4261 gactaccccc gtccgattct cggtggccgc gctcgcaggc cccgcctcgc cgaacatgtg
     4321 cgctgggacg cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat accagtgtgc
     4381 agatcttggc ccgcatttac aagactatct tgccagaaaa aaagcgtcgc agcaggtcat
     4441 caaaaatttt aaatggctag agacttatcg aaagcagcga gacaggcgcg aaggtgccac
     4501 cagattcgca cgcggcggcc ccagcgccca ggccaggcct caactcaagc acgaggcgaa
     4561 ggggctcctt aagcgcaagg cctcgaactc tcccacccac ttccaacccg aagctcggga
     4621 tcaagaatca cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg gccataaccc
     4681 gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct ggcggcaaac
     4741 ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc tcccccaccc
     4801 tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc gcgccacctt
     4861 ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac tgtgggcgat
     4921 gtgcgctctg cccactgacg ggcacatgca tggcggtaat acggttatcc acgcggccct
     4981 agagcgggcc cggtactaac caaactggat ctctgctgtc cctgtaataa acccgaaaat
     5041 tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt
     5101 ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga
     5161 tgaatactgc catttgtctg cagaattggc gcacgcagtg ccgatccgtt cactaatcga
     5221 atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt
     5281 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
     5341 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
     5401 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
     5461 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
     5521 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
     5581 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
     5641 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     5701 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     5761 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     5821 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     5881 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     5941 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     6001 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
     6061 agcggccggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc tctatcttgt
     6121 ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag agggttgcta
     6181 ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt tgtagctgtc
     6241 ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact gcgaatcgtt
     6301 ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt ccccctggcc
     6361 ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg acgctctcgc
     6421 acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc accagtcgcc
     6481 gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga gagagctcct
     6541 ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga ttttccacac
     6601 tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac
     6661 tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc
     6721 agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg caaaacgcgc
     6781 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     6841 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     6901 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     6961 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     7021 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     7081 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     7141 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     7201 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     7261 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     7321 taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta
     7381 catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca
     7441 agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac
     7501 aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     7561 accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa
     7621 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7681 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7741 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7801 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7861 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7921 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7981 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     8041 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     8101 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     8161 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8221 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8281 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8341 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8401 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     8461 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8521 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8581 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8641 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
