LOCUS       sequence-77194-e        8450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mouse Trific fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8450)
  AUTHORS   Short KM, Cox TC
  TITLE     Subclassification of the RBCC/TRIM superfamily reveals a novel motif
            necessary for microtubule binding.
  JOURNAL   J Biol Chem. 2006 Mar 31;281(13):8970-80. Epub 2006 Jan 23.
  PUBMED    16434393
REFERENCE   2  (bases 1 to 8450)
  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: "J Biol 
            Chem."; date: "2006-03-31"; volume: "281(13)"; pages: "8970-80. Epub
            2006 Jan 23"
COMMENT     Sequence Label: pcDNA-DEST53-mTrific
FEATURES             Location/Qualifiers
     source          1..8450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..63)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    52..276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      322..750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(409..428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     619..640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(759..779)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        764..1093
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      944..1079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1006..1025
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1160..1954
                     /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(1214..1233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1824..1843
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2128..2249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2165..2184)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2219..2238
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2298..2314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2298..2314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2311..2333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2322..2338
                     /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(2346..2352,2353..2370,2371..2376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2391..2412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2529..2546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2700..3288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2780..2799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3459..4250,4251..4319))
                     /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     4082..4101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4320..4424)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4499..4518)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4690..5069
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5070..5273
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5224..5244
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5318..5337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5318..5336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5360..5362,5363..5365,5366..6076)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(5393..5421)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6016..6036
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    6098..6122
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(8440..8450,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggtgataat taattaagat aaacccgctg atcagcctcg actgtgcctt
       61 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      121 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      181 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
      241 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
      301 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      361 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      541 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      601 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      661 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      721 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
      781 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      841 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      901 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
      961 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1021 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1081 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     1141 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     1201 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     1261 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     1321 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     1381 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     1441 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     1501 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     1561 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     1621 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     1681 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     1741 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     1801 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     1861 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     1921 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     1981 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     2041 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     2101 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     2161 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2221 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     2281 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2341 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2401 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2461 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2521 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2581 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2641 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2701 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2761 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2821 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2881 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2941 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3001 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3061 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3121 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3181 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3241 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3301 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3361 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3421 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3481 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3541 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3601 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3661 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3721 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3781 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3841 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3901 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3961 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4021 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4081 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4141 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4201 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4261 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4321 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4381 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4441 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     4501 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     4561 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     4621 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     4681 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     4741 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4801 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4861 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4921 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4981 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5041 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5101 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5161 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5221 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     5281 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     5341 cccaagctgg ctagacacca tggccagcaa aggagaagaa cttttcactg gagttgtccc
     5401 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
     5461 tgaaggtgat gctacatacg gaaagcttac ccttaaattt atttgcacta ctggaaaact
     5521 acctgttcca tggccaacac ttgtcactac tttctcttat ggtgttcaat gcttttcccg
     5581 ttatccggat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
     5641 acaggaacgc actatatctt tcaaagatga cgggaactac aagacgcgtg ctgaagtcaa
     5701 gtttgaaggt gatacccttg ttaatcgtat cgagttaaaa ggtattgatt ttaaagaaga
     5761 tggaaacatt ctcggacaca aactcgagta caactataac tcacacaatg tatacatcac
     5821 ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca acattgaaga
     5881 tggatccgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     5941 ccttttacca gacaaccatt acctgtcgac acaatctgcc ctttcgaaag atcccaacga
     6001 aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta cacatggcat
     6061 ggatgagctc tacaaaagcg gttggcctcc ggatcaaaca agtttgtaca aaaaagcagg
     6121 ctccgcggcc gcccccttca ccatgggtgg tgcccttgaa ataaatgcta ttaatccaga
     6181 gatggaggga tggagacaga caagcatgaa gaacatggag aaggaactgc tgtgcccagt
     6241 gtgccaagag atgtacaagc agccactagt gctgccttgt acccacaacg tgtgccaggc
     6301 ctgtgcccgg gaggttttgg gccagcaggg gtacataggc catggtgggg atccgagttc
     6361 ggagcccact tctcctgctt ccaccccttc tacccgcagc ccccgcctct ctcggagaac
     6421 tctccccaag ccagatcgct tggaccggct ccttaagtca ggctttggga cataccctgg
     6481 gcggaagcgt ggtgccttgc acccccagac gatcctattc ccgtgcccag cctgccaagg
     6541 cgatgtggag ctgggggaga ggggcctttc cgggcttttc cgaaacctga ccctagagcg
     6601 agtggtggag aggtaccgcc agagtgtgag tgtgggagga gccatcctgt gccagttgtg
     6661 caagccccca ccactagagg ccaccaaggg ctgcacagag tgtcgagcta ccttctgtaa
     6721 cgaatgcttc aagctcttcc acccctgggg cactcagaag gcgcagcatg agcccacgct
     6781 acccactctc tcattccgcc ccaagggtct gatgtgtcca gaccacaagg aagaggtgac
     6841 ccactactgt aagacatgtc aacgactggt gtgccaactc tgcagagtac ggcgtaccca
     6901 cagtgggcac aagatcacac cagtgctcag tgcctaccag gccctcaagg ataagctaac
     6961 aaagagcctg gcatacatct tgggaaatca ggacactgtg cagacccaga tttgtgagct
     7021 ggaagagacc atcaggcaca ctgaggtgag tggtcagcaa gcaaaagagg aggtgtccca
     7081 gctggttcgg ggactagggg ctgtgctgga agagaagcgg gcctcactgc ttcaggccat
     7141 tgaagaatgc cagcaagagc gcttatcccg gctcagcgcc cagatccatg aacaccagag
     7201 cctgctggat ggctcgggtc tggtgggtta tgcgcaggaa gtccttaagg aaacagacca
     7261 gccttgtttt gtacaagcag ccaaacagct gcataacagg attgcccgag ccactgaggc
     7321 cctccagaca ttccggccag ctgccagctc ctccttccgc cactgccagc tggatgtggg
     7381 gcgtgagatg aagctgttga ccgagcttag cttcctgaga gtgcccgagg cgcccgtcat
     7441 agacacccaa cgcacctttg cctatgatca aatcttcctg tgctggcggc tgccccctca
     7501 ctcaccacct gcctggcatt acaccgttga gttccggcgc acagatgtcc ccgcccagcc
     7561 aggccctaca cgctggcagc gccgggaaga ggtgaggggc accagcgccc ttcttgagaa
     7621 ccccgacacg ggctctgtgt acgtgctgcg tgtccgtggc tgcaacaagg ctggctacgg
     7681 agagtacagt gaagatgtgc acctgcacac acccccagcc cctgtcctgc acttcttcct
     7741 tgatggccgc tggggtgcga gccgagagcg gctagccatc agcaaggacc agcgagcggt
     7801 gcggagtatc ccagggcttc ccttgctgct ggctgctgag cggctgctca cgggctgcca
     7861 cctgagtgtg gacgtggtct tgggtgatgt ggctgtgacc cagggccgca gctactgggc
     7921 ctgtgcggta gaccctgcct cctacttggt caaggtgggc gtcgggctgg agagcaaact
     7981 tcaggaaagc ttccagggtg ccccggatgt gatcagcccc agatacgacc cggacagtgg
     8041 gcacgacagc ggtgccgagg acgctgccgt ggaggcgtta ccaccctttg ctttcctgac
     8101 catcggcatg ggcaagatcc tgctgggttc cggagcgagt tccaatgcgg ggctgacagg
     8161 gagggatggc cccaccacca gctgcacagt gcccctgcca ccccgcctag gtatctgttt
     8221 ggactacgag cgggggcggg tctccttcct ggatgccgtg tccttccgtg ggctgctcga
     8281 gtgccccctg gactgctcag ggcctgtgtg ccctgccttc tgcttcattg ggggtggcgc
     8341 tgttcagctg caggagcccg tgggcaccaa gcctgagagg aaagtcacca ttgggggctt
     8401 tgccaaactg gactgagaat tccaagggtg ggcgcgccga cccagctttc
//
