LOCUS       sequence-448505-        7910 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 7910)
  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 7910)
  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..7910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             341..352
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1992..2009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1998..2222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2268..2696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2355..2374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2565..2586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2705..2725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2710..3039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2890..3025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2952..2971
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3052..3651
                     /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(3052..3071)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3548..3568
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3825..3946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3862..3881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3916..3935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3995..4011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3995..4011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4008..4030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4019..4035
                     /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(4043..4049,4050..4067,4068..4073))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4088..4109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4226..4243)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4397..4985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4477..4496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5156..5947,5948..6016))
                     /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     5779..5798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6017..6121)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6196..6215)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6387..6766
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6767..6970
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6921..6941
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7015..7034
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7015..7033
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(7057..7059,7060..7062,7063..7773)
                     /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(7102..7123)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7363..7382)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7710..7731
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aattggcaag aaattcctcc ccagtgacat caatagtgga aaggtagaaa agctcgaagg
       61 tccatgtgtt ttgcaaattc aaaaaattcg caatgttgct gcaccaaagg ataatgaaga
      121 atctcaggct gcaccaagga tgctgcgatt acagatgact gatggtcata taagttgcac
      181 agcagtagaa tttagttata tgtcaaaaat aagcctgaac acaccacctg gaactaaagt
      241 taagctctca ggcattgttg acataaaaaa tggattcctg ctcttgaatg actctaacac
      301 cacagttctt ggtggtgaag tggaacacct tattgagaaa tgggagttac agagaagctt
      361 atcaaaacac aatagaagca atattggaac tgaaggtgga ccaccgcctt ttgtgccttt
      421 tggacagaag tgtgtatctc atgtccaagt ggatagcaga gaacttgatc gaagaaaaac
      481 attgcaagtt acaatgcctg tcaaacctac aaatgataat gatgaatttg aaaagcaaag
      541 gacggctgct attgctgaag ttgcaaagag caaggaagtt gatgagaaag ctctgaagca
      601 cataacggaa atgggcttca gtaaggaagc atcgaggcaa gctcttatgg ataatggcaa
      661 caacttagaa gcagcactga acgtacttct tacaagcaat aaacagaaac ctgttatggg
      721 tcctcctctg agaggtagag gaaaaggcag ggggcgaata agatctgaag atgaagagga
      781 cctgggaaat gcaaggccat cagcaccaag cacattattt gatttcttgg aatctaaaat
      841 gggaactttg aatgtggaag aacctaaatc acagccacag cagcttcatc agggacaata
      901 cagatcatca aatactgagc aaaatggagt aaaagataat aatcatctga gacatcctcc
      961 tcgaaatgat accaggcagc caagaaatga aaaaccgcct cgttttcaaa gagactccca
     1021 aaattcaaag tcagttttag aaggcagtgg attacctaga aatagaggtt ctgaaagacc
     1081 aagtacttct tcagtatctg aagtatgggc tgaagacaga atcaaatgtg atagaccgta
     1141 ttctagatat gacagaacta aagatacttc atatccttta ggttctcagc atagtgatgg
     1201 tgcttttaaa aaaagagata actctatgca aagcagatca ggaaaaggtc cctcctttgc
     1261 agaggcaaaa gaaaatccac ttcctcaagg atctgtagat tataataatc aaaaacgtgg
     1321 aaaaagagaa agccaaacat ctattcctga ttatttttat gacaggaaat cacaaacaat
     1381 aaataatgaa gctttcagtg gtataaaaat tgaaaaacat tttaatgtaa atactgatta
     1441 tcagaatcca gttcgaagta atagtttcat tggtgttcca aatggagaag tagaaatgcc
     1501 actgaaagga agacgaatag gacctattaa gccagcagga cctgtcacag ctgtaccctg
     1561 tgatgataaa atattttaca atagtgggcc caaacgaaga tctgggccaa ttaagccaga
     1621 aaaaatacta gaatcatcta ttcctatgga gtatgcaaaa atgtggaaac ctggagatga
     1681 atgttttgca ctttattggg aagacaacaa gttttaccgg gcagaagttg aagccctcca
     1741 ttcttcgggt atgacagcag ttgttaaatt cattgactac ggaaactatg aagaggtgct
     1801 actgagcaat atcaagccca ttcaaacaga ggcatgggag gaagaaggca cctacgatca
     1861 aactctggag ttccgtaggg gaggtgatgg ccagccaaga cgatccactc ggccaaccca
     1921 acagttttac caaccacccc gggctcggaa ctaactcgag tctagagggc ccgtttaaac
     1981 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2041 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2101 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2161 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2221 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2281 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2341 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2401 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2461 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2521 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2581 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2641 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2701 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2761 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2821 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2881 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2941 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3001 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccaac catgaccgag
     3061 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3121 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3181 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3241 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3301 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3361 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3421 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3481 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3541 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3601 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agcgggactc
     3661 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     3721 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     3781 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     3841 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3901 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     3961 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4021 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4081 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4141 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4201 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4261 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4321 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4381 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4441 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4501 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4561 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4621 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4681 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4741 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4801 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4861 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4921 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4981 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5041 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5101 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5161 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5221 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5281 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5341 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5401 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5461 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5521 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5581 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5641 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5701 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5761 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5821 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5881 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5941 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6001 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6121 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     6181 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     6241 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     6301 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     6361 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     6421 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     6481 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     6541 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     6601 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     6661 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     6721 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     6781 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     6841 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     6901 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     6961 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     7021 gactcactat agggagaccc aagctggcta gcaaccatgg tgagcaaggg cgaggagctg
     7081 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     7141 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     7201 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     7261 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     7321 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     7381 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     7441 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     7501 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     7561 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     7621 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     7681 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     7741 gggatcactc tcggcatgga cgagctgtac aaggcccagg tggccggcgc ggcgttgtcc
     7801 caggcgggtt ggtatctttc agatgaaggc attgaagctt gcacaagctc tccagacaaa
     7861 gtcaatgtaa atgacatcat cctgattgct ctcaatacag atctgagaac
//
