LOCUS       sequence-385266-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_494-546.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  AUTHORS   Calero-Cuenca FJ, Osorio DS, Carvalho-Marques S, Sridhara SC, 
            Oliveira LM, Jiao Y, Diaz J, Janota CS, Cadot B, Gomes ER
  TITLE     Ctdnep1 and Eps8L2 regulate dorsal actin cables for nuclear 
            positioning during cell migration.
  JOURNAL   Curr Biol. 2021 Apr 12;31(7):1521-1530.e8. doi: 
            10.1016/j.cub.2021.01.007. Epub 2021 Feb 9.
  PUBMED    33567288
REFERENCE   2  (bases 1 to 5529)
  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.cub.2021.01.007"; journalName: "Curr Biol"; date: 
            "2021-04-12- 12"; volume: "31"; issue: "7"; pages: "1521-1530.e8"
COMMENT     Sequence Label: pDEST15-Eps8L2_494-546
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      133..721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     622..641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     875..892
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    907..1046
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(972..991)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1132..1154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1148..1339)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2801..2820
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2904..2923
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2904..2922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2954..2976
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2984..3637
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    3671..3695
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3870..3894)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3958..3976)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3972..4019
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4385..4390,4391..4407,4408..4413))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4440..4458)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4526..4630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4631..4699,4700..5491)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4849..4868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
       61 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
      121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
      181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
      241 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
      301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      781 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      841 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      901 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
      961 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     1021 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     1081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     1201 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     1261 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     1321 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     1381 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     1441 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     1501 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     1561 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     1621 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     1681 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     1741 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     1801 cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg
     1861 acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt
     1921 gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca
     1981 ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag
     2041 ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc
     2101 cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg
     2161 aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat
     2221 cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc
     2281 gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt
     2341 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     2401 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     2461 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     2521 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     2581 ggctctcaag ggcatcggtc gatcgacgct ctcccttatg cgactcctgc attaggaagc
     2641 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     2701 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     2761 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     2821 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     2881 tcgagatctc gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc
     2941 ctctagaaat aattttgttt aactttaaga aggagatata catatgtccc ctatactagg
     3001 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
     3061 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
     3121 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
     3181 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
     3241 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
     3301 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
     3361 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
     3421 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
     3481 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
     3541 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
     3601 gcaagccacg tttggtggtg gcgaccatcc tccaaaatcg gatctggttc cgcgtccatg
     3661 gtcgaatcaa acaagtttgt acaaaaaagc aggctccatg ccagccatgg ccaaatatgt
     3721 gaaaattctc tatgacttca cagcccgaaa tgccaacgag ctatcggtgc tcaaggatga
     3781 ggtcctagag gtgctggagg acggccggca gtggtggaag ctgcgcagcc gcagcggcca
     3841 ggcggggtac gtgccctgca acatcctaga cccagctttc ttgtacaaag tggtttgatt
     3901 cgacccggga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     3961 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     4021 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
     4081 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
     4141 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
     4201 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
     4261 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     4321 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     4381 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg
     4441 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4621 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4681 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4741 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4801 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4861 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4921 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4981 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5041 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5101 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5161 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5221 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5281 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5341 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5401 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5461 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5521 ttagattga
//
