LOCUS       sequence-178622-        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human TRIM32 fused to c-terminal EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  AUTHORS   Fu B, Wang L, Ding H, Schwamborn JC, Li S, Dorf ME
  TITLE     TRIM32 Senses and Restricts Influenza A Virus by Ubiquitination of 
            PB1 Polymerase.
  JOURNAL   PLoS Pathog. 2015 Jun 9;11(6):e1004960. doi: 
            10.1371/journal.ppat.1004960. eCollection 2015 Jun.
  PUBMED    26057645
REFERENCE   2  (bases 1 to 6659)
  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: "PLoS 
            Pathog."; date: "2015-06-9"; volume: "11(6):e1004960. doi"; pages: "
            10.1371/journal.ppat.1004960. eCollection 2015 Jun"
COMMENT     Sequence Label: pEGFP-N1_hTRIM32
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      619..628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2599..2608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2605..2607,2608..2610,2611..3324)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2605..2607,2608..2610,2611..3321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2650..2671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2911..2930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3258..3279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3447..3568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3484..3503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3538..3557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3575..4030)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3712..3733)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3924..3943
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4057..4161
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4163..4520
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4163..4354
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4186..4206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4371..4506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4555..5349
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4555..5349
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4609..4628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5219..5238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5537..5556)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5581..5628
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5957..6545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6446..6465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatggct gcagcagcag cttctcacct gaacctggat
      661 gccctccggg aagtgctaga atgccccatc tgcatggagt ccttcacaga agagcagctg
      721 cgtcccaagc ttctgcactg tggccatacc atctgccgcc agtgcctgga gaagctattg
      781 gccagtagca tcaatggtgt ccgctgtccc ttttgcagca agattacccg cataaccagc
      841 ttgacccagc tgacagacaa tctgacagtg ctaaagatca ttgatacagc tgggctcagc
      901 gaggctgtgg ggctgctcat gtgtcggtcc tgtgggcggc gtctgccccg gcaattctgc
      961 cggagctgtg gtttggtgtt atgtgagccc tgccgggagg cagaccatca gcctcctggc
     1021 cactgtacac tccctgtcaa agaagcagct gaggagcggc gtcgggactt tggagagaag
     1081 ttaactcgtc tgcgggaact tatgggggag ctgcagcggc ggaaggcagc cttggaaggt
     1141 gtctccaagg accttcaggc aaggtataaa gcagttctcc aggagtatgg gcatgaggag
     1201 cgcagggtcc aggatgagct ggctcgctct cggaagttct tcacaggctc tttggctgaa
     1261 gttgagaagt ccaatagtca agtggtagag gagcagagtt acctgcttaa cattgcagag
     1321 gtgcaggctg tgtctcgctg tgactacttc ctggccaaga tcaagcaggc agatgtagca
     1381 ctactggagg agacagctga tgaggaggag ccagagctca ctgccagctt gcctcgggag
     1441 ctcaccctgc aagatgtgga gctccttaag gtaggtcatg ttggccccct ccaaattgga
     1501 caagctgtta agaagccccg gacagttaac gtggaagatt cctgggccat ggaggccaca
     1561 gcgtctgctg cctctacctc tgttactttt agagagatgg acatgagccc ggaggaagtg
     1621 gttgccagcc ctagggcctc acctgctaaa cagcggggtc ctgaggcagc ctccaatatc
     1681 cagcagtgcc tctttctcaa gaagatgggg gccaaaggca gcactccagg aatgttcaat
     1741 cttccagtca gtctctacgt gaccagtcaa ggtgaagtac tagtcgctga ccgtggtaac
     1801 tatcgtatac aagtctttac ccgcaaaggc tttttgaagg aaatccgccg cagccccagt
     1861 ggcattgata gctttgtgct aagcttcctt ggggcagatc tacccaacct cactcctctc
     1921 tcagtggcaa tgaactgcca ggggctgatt ggtgtgactg acagctatga taactccctc
     1981 aaggtatata ccttggatgg ccactgcgtg gcctgtcaca ggagccagct gagcaaacca
     2041 tggggtatca cagccttgcc atctggccag tttgtagtaa ccgatgtgga aggtggaaag
     2101 ctttggtgtt tcacagttga tcgaggatca ggggtggtca aatacagctg cctatgtagt
     2161 gctgtgcggc ccaaatttgt cacctgtgat gctgagggca ccgtctactt cacccagggc
     2221 ttaggcctca atctggagaa tcggcagaat gagcaccacc tggagggtgg cttttccatt
     2281 ggctctgtag gccctgatgg gcagctgggt cgccagatta gccacttctt ctcggagaat
     2341 gaggatttcc gctgcattgc tggcatgtgt gtggatgctc gtggtgatct catcgtggct
     2401 gacagtagtc gcaaggaaat tctccatttt cctaagggtg ggggctatag tgtccttatt
     2461 cgagagggac ttacctgtcc ggtgggcata gccctaactc ctaaggggca gctgctggtc
     2521 ttggactgtt gggatcattg catcaagatc tacagctacc atctgagaag atattccacc
     2581 ccacgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2641 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2701 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2761 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2821 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2881 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2941 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3001 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3061 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3121 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3181 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3241 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3301 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
     3361 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3421 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3541 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     3601 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3661 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3721 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3781 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3841 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3901 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3961 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4021 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4081 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4141 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4201 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4261 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4381 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4441 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4501 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4561 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4621 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4681 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4741 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4801 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4861 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4921 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4981 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5041 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5101 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5161 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5221 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5281 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5341 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5401 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5461 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5521 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5581 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5641 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5701 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5761 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5821 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6601 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
