LOCUS       sequence-386573-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_544-715.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5877)
  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 5877)
  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_544-715
FEATURES             Location/Qualifiers
     source          1..5877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1278..1297
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1381..1400
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1381..1399
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1431..1453
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1461..2114
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    2148..2172
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2695..2719)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2783..2801)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2797..2844
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3210..3215,3216..3232,3233..3238))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3265..3283)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3351..3455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3456..3524,3525..4316)
                     /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(3674..3693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4487..5075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4976..4995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5229..5246
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5261..5400
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5326..5345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5486..5508
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5502..5693)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
       61 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
      121 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
      181 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
      241 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
      301 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
      361 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
      421 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
      481 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
      541 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga
      601 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
      661 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
      721 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
      781 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
      841 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
      901 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
      961 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     1021 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgat
     1081 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     1141 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     1201 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     1261 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     1321 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
     1381 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
     1441 tttaagaagg agatatacat atgtccccta tactaggtta ttggaaaatt aagggccttg
     1501 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
     1561 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
     1621 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
     1681 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
     1741 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
     1801 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
     1861 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca
     1921 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
     1981 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
     2041 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
     2101 accatcctcc aaaatcggat ctggttccgc gtccatggtc gaatcaaaca agtttgtaca
     2161 aaaaagcagg ctccatgaac atcctaggcg aggcgcgacc ggaggacgcc ggcgccccgt
     2221 tcgagcaggc cggtcagaag tactggggcc ccgccagccc gacccacaag ctacccccaa
     2281 gcttcccggg caataaggac gaactcatgc agcacatgga cgaggtcaac gacgagctca
     2341 tccgcaagat atcgaatata agggcgcagc cacagaggca cttccgcgtg gagcgcagcc
     2401 agcccgtgag ccagccgctc acctacgagt cgggtccgga cgaggtccgc gcctggctgg
     2461 aagccaaggc cttcagcccg cggatcgtgg agaacctggg catcctgacc gggccgcagc
     2521 tcttctccct caacaaggag gagctgaaga aagtgtgcgg cgaggagggc gtccgcgtgt
     2581 acagccagct caccatgcag aaggccttcc tggagaagca gcaaagtggg tcggagctgg
     2641 aagaactcat gaacaagttt cattccatga atcagaggag gggggaggac agcgacccag
     2701 ctttcttgta caaagtggtt tgattcgacc cgggatccgg ctgctaacaa agcccgaaag
     2761 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2821 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tccacaggac
     2881 gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac
     2941 tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat
     3001 caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat
     3061 atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt
     3121 gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg
     3181 ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac
     3241 atgagaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3301 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3361 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3421 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3481 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3541 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3601 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3661 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     3721 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3781 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3841 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3901 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3961 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     4021 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4081 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4141 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4201 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4261 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4321 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4621 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5161 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5221 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5281 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     5341 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     5401 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5461 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5521 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     5581 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     5641 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     5701 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     5761 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     5821 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccag
//
