LOCUS       sequence-346203-        8125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRIM46 tagged with GFP .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8125)
  AUTHORS   van Beuningen SFB, Will L, Harterink M, Chazeau A, van Battum EY, 
            Frias CP, Franker MAM, Katrukha EA, Stucchi R, Vocking K, Antunes 
            AT, Slenders L, Doulkeridou S, Sillevis Smitt P, Altelaar AFM, Post 
            JA, Akhmanova A, Pasterkamp RJ, Kapitein LC, de Graaff E, Hoogenraad
            CC
  TITLE     TRIM46 Controls Neuronal Polarity and Axon Specification by Driving 
            the Formation of Parallel Microtubule Arrays.
  JOURNAL   Neuron. 2015 Dec 16;88(6):1208-1226. doi: 
            10.1016/j.neuron.2015.11.012. Epub 2015 Dec 6.
  PUBMED    26671463
REFERENCE   2  (bases 1 to 8125)
  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.neuron.2015.11.012"; journalName: "Neuron"; date: 
            "2015-12-16- 16"; volume: "88"; issue: "6"; pages: "1208-1226"
COMMENT     Sequence Label: GFP-Trim46
FEATURES             Location/Qualifiers
     source          1..8125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          340..1166
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     regulatory      1190..1199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1196..1198,1199..1201,1202..1912)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1196..1198,1199..1201,1202..1912)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1241..1262)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1502..1521)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1849..1870
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4351..4485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4388..4407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4442..4461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4512..4528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4512..4528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4525..4547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4536..4552
                     /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(4560..4566,4567..4584,4585..4590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4742..4759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4913..5501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4993..5012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5672..6463,6464..6532))
                     /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     6295..6314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     6720..6738
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6776..6798)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6898..6917
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7149..7171
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7163..7180
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7164..7180
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        7746..8125
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
      301 cggtgcattg gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc
      361 tataggccca cccccttggc ttcttatgca tgctatactg tttttggctt ggggtctata
      421 cacccccgct tcctcatgtt ataggtgatg gtatagctta gcctataggt gtgggttatt
      481 gaccattatt gaccactccc ctattggtga cgatactttc cattactaat ccataacatg
      541 gctctttgcc acaactctct ttattggcta tatgccaata cactgtcctt cagagactga
      601 cacggactct gtatttttac aggatggggt ctcatttatt atttacaaat tcacatatac
      661 aacaccaccg tccccagtgc ccgcagtttt tattaaacat aacgtgggat ctccacgcga
      721 atctcgggta cgtgttccgg acatgggctc ttctccggta gcggcggagc ttctacatcc
      781 gagccctgct cccatgcctc cagcgactca tggtcgctcg gcagctcctt gctcctaaca
      841 gtggaggcca gacttaggca cagcacgatg cccaccacca ccagtgtgcc gcacaaggcc
      901 gtggcggtag ggtatgtgtc tgaaaatgag ctcggggagc gggcttgcac cgctgacgca
      961 tttggaagac ttaaggcagc ggcagaagaa gatgcaggca gctgagttgt tgtgttctga
     1021 taagagtcag aggtaactcc cgttgcggtg ctgttaacgg tggagggcag tgtagtctga
     1081 gcagtactcg ttgctgccgc gcgcgccacc agacataata gctgacagac taacagactg
     1141 ttcctttcca tgggtctttt ctgcagtcac cgtccttgac acgaagcttg ccaccatggt
     1201 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1261 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1321 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1381 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1441 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1501 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1561 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1621 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1681 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1741 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1801 gagcacccag tctgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1861 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtccggagg
     1921 cgcgcctgaa ttcatggctg agggtgagga tatgcagacc ttcacttcca tcatggatgc
     1981 actagtccgc atcagtacaa gcatgaagaa catggagaag gaactgctgt gcccagtgtg
     2041 ccaagagatg tacaagcagc cactagtgct gccttgtacc cacaacgtgt gccaggcctg
     2101 tgcccgggag gttttgggcc agcaggggta cataggccat ggtggggatc cgagttcgga
     2161 gcccacttct cctgcttcca ccccttctac ccgcagcccc cgcctctctc ggagaactct
     2221 ccccaagcca gatcgcttgg accggctcct taagtcaggc tttgggacat accctgggcg
     2281 gaagcgtggt gccttgcacc cccagacgat cctattcccg tgcccagcct gccaaggcga
     2341 tgtggagctg ggggagaggg gcctttccgg gcttttccga aacctgaccc tagagcgagt
     2401 ggtggagagg taccgccaga gtgtgagtgt gggaggagcc atcctgtgcc agttgtgcaa
     2461 gcccccacca ctagaggcca ccaagggctg cacagagtgt cgagctacct tctgtaacga
     2521 atgcttcaag ctcttccacc cctggggcac tcagaaggcg cagcatgagc ccacgctacc
     2581 cactctctca ttccgcccca agggtctgat gtgtccagac cacaaggaag aggtgaccca
     2641 ctactgtaag acatgtcaac gactggtgtg ccaactctgc agagtacggc gtacccacag
     2701 tgggcacaag atcacaccag tgctcagtgc ctaccaggcc ctcaaggata agctaacaaa
     2761 gagcctggca tacatcttgg gaaatcagga cactgtgcag acccagattt gtgagctgga
     2821 agagaccatc aggcacactg aggtgagtgg tcagcaagca aaagaggagg tgtcccagct
     2881 ggttcgggga ctaggggctg tgctggaaga gaagcgggcc tcactgcttc aggccattga
     2941 agaatgccag caagagcgct tatcccggct cagcgcccag atccatgaac accagagcct
     3001 gctggatggc tcgggtctgg tgggttatgc gcaggaagtc cttaaggaaa cagaccagcc
     3061 ttgttttgta caagcagcca aacagctgca taacaggatt gcccgagcca ctgaggccct
     3121 ccagacattc cggccagctg ccagctcctc cttccgccac tgccagctgg atgtggggcg
     3181 tgagatgaag ctgttgaccg agcttagctt cctgagagtg cccgaggcgc ccgtcataga
     3241 cacccaacgc acctttgcct atgatcaaat cttcctgtgc tggcggctgc cccctcactc
     3301 accacctgcc tggcattaca ccgttgagtt ccggcgcaca gatgtccccg cccagccagg
     3361 ccctacacgc tggcagcgcc gggaagaggt gaggggcacc agcgcccttc ttgagaaccc
     3421 cgacacgggc tctgtgtacg tgctgcgtgt ccgtggctgc aacaaggctg gctacggaga
     3481 gtacagtgaa gatgtgcacc tgcacacacc cccagcccct gtcctgcact tcttccttga
     3541 tggccgctgg ggtgcgagcc gagagcggct agccatcagc aaggaccagc gagcggtgcg
     3601 gagtatccca gggcttccct tgctgctggc tgctgagcgg ctgctcacgg gctgccacct
     3661 gagtgtggac gtggtcttgg gtgatgtggc tgtgacccag ggccgcagct actgggcctg
     3721 tgcggtagac cctgcctcct acttggtcaa ggtgggcgtc gggctggaga gcaaacttca
     3781 ggaaagcttc cagggtgccc cggatgtgat cagccccaga tacgacccgg acagtgggca
     3841 cgacagcggt gccgaggacg ctgccgtgga ggcgttacca ccctttgctt tcctgaccat
     3901 cggcatgggc aagatcctgc tgggttccgg agcgagttcc aatgcggggc tgacagggag
     3961 ggatggcccc accaccagct gcacagtgcc cctgccaccc cgcctaggta tctgtttgga
     4021 ctacgagcgg gggcgggtct ccttcctgga tgccgtgtcc ttccgtgggc tgctcgagtg
     4081 ccccctggac tgctcagggc ctgtgtgccc tgccttctgc ttcattgggg gtggcgctgt
     4141 tcagctgcag gagcccgtgg gcaccaagcc tgagaggaaa gtcaccattg ggggctttgc
     4201 caaactggac tgagtcgact gcggccgcct ctagataatt cattgatcat aatcagccat
     4261 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4321 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4381 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4441 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgatcc gtaccgagct
     4501 cgcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4561 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaacta
     4621 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4681 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4741 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4801 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4861 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4921 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4981 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5041 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5101 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5161 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5221 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5281 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5341 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5401 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5461 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5521 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5581 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5641 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5701 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5761 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5821 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5881 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5941 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6001 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6061 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6121 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6181 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6241 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6301 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6361 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6421 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6481 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6541 tttttcaata agcggcctcg agttattatt gaagcattta tcagggttat tgtctcatga
     6601 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6661 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6721 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6781 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6841 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6901 catcagagca gattgtactg agagtgcacc atatgaagac gtcgcctcct cactacttct
     6961 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagg
     7021 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7081 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7141 agggttttcc cagtcacgac gttgtaaaac gacggccagt gcccgtgtca aggacggtga
     7201 ctgcagtgaa taataaaatg tgtgtttgtc cgaaatacgc gttttgagat ttctgtcgcc
     7261 gactaaattc atgtcgcgcg atagtggtgt ttatcgccga tagagatggc gatattggaa
     7321 aaatcgatat ttgaaaatat ggcatattga aaatgtcgcc gatgtgagtt tctgtgtaac
     7381 tgatatcgcc atttttccaa aagtgatttt tgggcatacg cgatatctgg cgatagcgct
     7441 tatatcgttt acgggggatg gcgatagacg actttggtga cttgggcgat tctgtgtgtc
     7501 gcaaatatcg cagtttcgat ataggtgaca gacgatatga ggctatatcg ccgatagagg
     7561 cgacatcaag ctggcacatg gccaatgcat atcgatctat acattgaatc aatattggcc
     7621 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
     7681 tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc
     7741 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     7801 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7861 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7921 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7981 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     8041 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     8101 cgtattagtc atcgctatta ccatg
//
