LOCUS       sequence-441714-        8048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mEGFP-fusion protein of TDRD3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8048)
  AUTHORS   Chen X, Fansler MM, Janjos U, Ule J, Mayr C
  TITLE     The FXR1 network acts as signaling scaffold for actomyosin 
            remodeling.
  JOURNAL   Cell. 5 August 2024. doi: 10.1016/j.cell.2024.07.015.
  PUBMED    37961296
REFERENCE   2  (bases 1 to 8048)
  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: "Cell. 5 
            August 2024. doi: 10.1016/j.cell.2024.07.015."
COMMENT     Sequence Label: pcDNA3puro-mGFP-TDRD3
FEATURES             Location/Qualifiers
     source          1..8048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(905..907,908..910,911..1621)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(950..971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1211..1230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2099..2110
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3891..3908)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3897..4121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4167..4595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4254..4273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4464..4485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4604..4624)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4609..4938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4789..4924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4851..4870
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4951..5550
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4951..4970)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5447..5467
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5724..5845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5761..5780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5815..5834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5894..5910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5894..5910)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5907..5929)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5918..5934
                     /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(5942..5948,5949..5966,5967..5972))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5987..6008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6125..6142)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6296..6881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6376..6395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7052..7843,7844..7912))
                     /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     7675..7694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7913..8017)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aaccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      961 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1021 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1081 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1141 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1201 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1261 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1321 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1381 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1501 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 ggcccaggtg gccggcgcgg cgttgtccca ggcgggttgg tatctttcag atgaaggcat
     1681 tgaagcttgc acaagctctc cagacaaagt caatgtaaat gacatcatcc tgattgctct
     1741 caatacagat ctgagaacaa ttggcaagaa attcctcccc agtgacatca atagtggaaa
     1801 ggtagaaaag ctcgaaggtc catgtgtttt gcaaattcaa aaaattcgca atgttgctgc
     1861 accaaaggat aatgaagaat ctcaggctgc accaaggatg ctgcgattac agatgactga
     1921 tggtcatata agttgcacag cagtagaatt tagttatatg tcaaaaataa gcctgaacac
     1981 accacctgga actaaagtta agctctcagg cattgttgac ataaaaaatg gattcctgct
     2041 cttgaatgac tctaacacca cagttcttgg tggtgaagtg gaacacctta ttgagaaatg
     2101 ggagttacag agaagcttat caaaacacaa tagaagcaat attggaactg aaggtggacc
     2161 accgcctttt gtgccttttg gacagaagtg tgtatctcat gtccaagtgg atagcagaga
     2221 acttgatcga agaaaaacat tgcaagttac aatgcctgtc aaacctacaa atgataatga
     2281 tgaatttgaa aagcaaagga cggctgctat tgctgaagtt gcaaagagca aggaaaccaa
     2341 gacatttgga ggaggtggtg gtggtgctag aagtaatctc aatatgaatg ctgctggtaa
     2401 ccgaaatagg gaagttttac agaaagaaaa gtcaaccaaa tcagagggaa aacatgaagg
     2461 tgtctataga gaactggttg atgagaaagc tctgaagcac ataacggaaa tgggcttcag
     2521 taaggaagca tcgaggcaag ctcttatgga taatggcaac aacttagaag cagcactgaa
     2581 cgtacttctt acaagcaata aacagaaacc tgttatgggt cctcctctga gaggtagagg
     2641 aaaaggcagg gggcgaataa gatctgaaga tgaagaggac ctgggaaatg caaggccatc
     2701 agcaccaagc acattatttg atttcttgga atctaaaatg ggaactttga atgtggaaga
     2761 acctaaatca cagccacagc agcttcatca gggacaatac agatcatcaa atactgagca
     2821 aaatggagta aaagataata atcatctgag acatcctcct cgaaatgata ccaggcagcc
     2881 aagaaatgaa aaaccgcctc gttttcaaag agactcccaa aattcaaagt cagttttaga
     2941 aggcagtgga ttacctagaa atagaggttc tgaaagacca agtacttctt cagtatctga
     3001 agtatgggct gaagacagaa tcaaatgtga tagaccgtat tctagatatg acagaactaa
     3061 agatacttca tatcctttag gttctcagca tagtgatggt gcttttaaaa aaagagataa
     3121 ctctatgcaa agcagatcag gaaaaggtcc ctcctttgca gaggcaaaag aaaatccact
     3181 tcctcaagga tctgtagatt ataataatca aaaacgtgga aaaagagaaa gccaaacatc
     3241 tattcctgat tatttttatg acaggaaatc acaaacaata aataatgaag ctttcagtgg
     3301 tataaaaatt gaaaaacatt ttaatgtaaa tactgattat cagaatccag ttcgaagtaa
     3361 tagtttcatt ggtgttccaa atggagaagt agaaatgcca ctgaaaggaa gacgaatagg
     3421 acctattaag ccagcaggac ctgtcacagc tgtaccctgt gatgataaaa tattttacaa
     3481 tagtgggccc aaacgaagat ctgggccaat taagccagaa aaaatactag aatcatctat
     3541 tcctatggag tatgcaaaaa tgtggaaacc tggagatgaa tgttttgcac tttattggga
     3601 agacaacaag ttttaccggg cagaagttga agccctccat tcttcgggta tgacagcagt
     3661 tgttaaattc attgactacg gaaactatga agaggtgcta ctgagcaata tcaagcccat
     3721 tcaaacagag gcatgggagg aagaaggcac ctacgatcaa actctggagt tccgtagggg
     3781 aggtgatggc cagccaagac gatccactcg gccaacccaa cagttttacc aaccaccccg
     3841 ggctcggaac taactcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3901 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3961 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4021 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4081 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4141 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4201 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4261 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4321 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4381 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4441 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4501 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4561 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4621 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4681 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4741 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4801 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4861 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4921 ggcctaggct tttgcaaaaa gctcccaacc atgaccgagt acaagcccac ggtgcgcctc
     4981 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5041 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5101 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5161 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5221 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5281 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5341 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5401 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5461 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5521 tggtgcatga cccgcaagcc cggtgcctga gcgggactct ggggttcgaa atgaccgacc
     5581 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5641 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5701 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5821 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5881 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5941 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6001 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6721 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     6841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6961 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7021 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7081 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7141 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7201 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7261 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7321 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7381 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7441 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7501 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7561 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7621 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7681 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7741 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7801 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7861 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8041 ctgacgtc
//
