LOCUS       sequence-386577-        5910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_1-183.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5910)
  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 5910)
  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_1-183
FEATURES             Location/Qualifiers
     source          1..5910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    239..378
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(304..323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     464..486
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(480..671)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2133..2152
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2236..2255
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2236..2254
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2286..2308
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2316..2969
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    3003..3027
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3583..3607)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3671..3689)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3685..3732
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4098..4103,4104..4120,4121..4126))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4153..4171)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4239..4343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4344..4412,4413..5204)
                     /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(4562..4581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5375..5910,1..53)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5864..5883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
       61 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      121 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      181 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      241 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
      301 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
      361 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      421 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
      481 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
      541 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
      601 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
      661 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
      721 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
      781 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
      841 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
      901 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
      961 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     1021 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     1081 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     1141 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     1201 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     1261 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg
     1321 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc
     1381 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga
     1441 tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc
     1501 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc
     1561 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca
     1621 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga
     1681 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     1741 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     1801 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     1861 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     1921 agggcatcgg tcgatcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     1981 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     2041 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     2101 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     2161 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     2221 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
     2281 ataattttgt ttaactttaa gaaggagata tacatatgtc ccctatacta ggttattgga
     2341 aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg
     2401 aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg
     2461 gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta
     2521 tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc
     2581 gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa
     2641 ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa
     2701 tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa
     2761 cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt
     2821 gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa
     2881 ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca
     2941 cgtttggtgg tggcgaccat cctccaaaat cggatctggt tccgcgtcca tggtcgaatc
     3001 aaacaagttt gtacaaaaaa gcaggctcca tgatgagcca gtccggggcc gtgagctgct
     3061 gcccgggtgc caccaatggc agcctgggcc ggtccgacgg tgtggccaag atgagcccca
     3121 aggacctgtt tgagcagagg aagaagtatt ccaactccaa cgtcatcatg cacgagacct
     3181 cgcagtacca cgtccagcac ctggccacat tcatcatgga caagagcgaa gccatcacgt
     3241 ctgtggacga cgccatccgg aagctggtgc agctgagctc caaggagaag atctggaccc
     3301 aggagatgct gctgcaggtg aacgaccagt cgctgcggct gctggacatc gagtcacagg
     3361 aggagctgga agacttcccg ctgcccacgg tgcagcgcag ccagacggtc ctcaaccagc
     3421 tgcgctaccc gtctgtgctg ctgctcgtgt gccaggactc ggagcagagc aagccggatg
     3481 tccacttctt ccactgcgat gaggtggagg cagagctggt gcacgaggac atcgagagcg
     3541 cgttggccga ctgccggctg ggcaagaaga tgcggccgca ggacccagct ttcttgtaca
     3601 aagtggtttg attcgacccg ggatccggct gctaacaaag cccgaaagga agctgagttg
     3661 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     3721 aggggttttt tgctgaaagg aggaactata tccggatatc cacaggacgg gtgtggtcgc
     3781 catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc
     3841 aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag
     3901 cgctagcagc acgccatagt gactggcgat gctgtcggaa tggacgatat cccgcaagag
     3961 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
     4021 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
     4081 ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattctt
     4141 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4201 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4261 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4321 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4381 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4441 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4501 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4561 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4621 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4681 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4741 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4801 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4861 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     4921 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4981 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5041 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5101 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5161 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5221 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5281 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5341 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5401 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5461 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5521 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5581 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5641 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5701 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5761 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5821 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5881 tttatagtcc tgtcgggttt cgccacctct
//
