LOCUS       sequence-385272-        5583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_297-370.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5583)
  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 5583)
  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_297-370
FEATURES             Location/Qualifiers
     source          1..5583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     356..373
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    388..527
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(453..472)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     613..635
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(629..820)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2282..2301
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2385..2404
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2385..2403
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2435..2457
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2465..3118
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    3152..3176
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3405..3429)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3493..3511)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3507..3554
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3920..3925,3926..3942,3943..3948))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3975..3993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4061..4165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4166..4234,4235..5026)
                     /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(4384..4403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5197..5583,1..202)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
       61 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      241 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      301 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
      361 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
      421 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      481 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
      541 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
      601 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
      661 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
      721 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
      781 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
      841 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
      901 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
      961 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     1021 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     1081 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     1141 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     1201 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     1261 acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag atgcgccgcg
     1321 tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat
     1381 tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga
     1441 ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc
     1501 ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct
     1561 cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag ttaggctggt
     1621 aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag
     1681 catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa
     1741 ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat
     1801 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     1861 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     1921 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     1981 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     2041 gaaggagctg actgggttga aggctctcaa gggcatcggt cgatcgacgc tctcccttat
     2101 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     2161 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     2221 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     2281 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     2341 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     2401 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
     2461 acatatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac ccactcgact
     2521 tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg atgaaggtga
     2581 taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc cttattatat
     2641 tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag ctgacaagca
     2701 caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg aaggagcggt
     2761 tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg aaactctcaa
     2821 agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca
     2881 taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt atgacgctct
     2941 tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat tagtttgttt
     3001 taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca gcaagtatat
     3061 agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc
     3121 ggatctggtt ccgcgtccat ggtcgaatca aacaagtttg tacaaaaaag caggctccat
     3181 gccagcagag ggcgtcctca cactgcgggc acggcccccc tctgagggcg agttcatcga
     3241 ctgcttccag aaaatcaagc tggcgattaa cttgctggca aagctgcaga agcacatcca
     3301 gaaccccagc gccgcggagc tcgtgcactt cctcttcggg cctctggacc tgatcgtcaa
     3361 cacctgcagt ggcccagaca tcgcacgctc cgtctcctgc ccagacccag ctttcttgta
     3421 caaagtggtt tgattcgacc cgggatccgg ctgctaacaa agcccgaaag gaagctgagt
     3481 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3541 tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac gggtgtggtc
     3601 gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac tgggcggcgg
     3661 ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat caacgcatat
     3721 agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat atcccgcaag
     3781 aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt gacggtgccg
     3841 aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg ttagcaattt
     3901 aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac atgagaattc
     3961 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4021 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4081 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4141 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4201 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4261 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4321 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4381 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     4441 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4501 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4561 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4621 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4681 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     4741 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4801 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4861 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4921 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4981 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5041 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5101 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5341 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5461 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5521 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5581 gcg
//
