LOCUS       sequence-386575-        5718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_181-299.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5718)
  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 5718)
  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_181-299
FEATURES             Location/Qualifiers
     source          1..5718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      135..723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     624..643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     877..894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    909..1048
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(974..993)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1134..1156
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1150..1341)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2803..2822
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2906..2925
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2906..2924
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2956..2978
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2986..3639
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    3673..3697
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4061..4085)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4149..4167)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4163..4210
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4576..4581,4582..4598,4599..4604))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4631..4649)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4717..4821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4822..4890,4891..5682)
                     /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(5040..5059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
       61 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
      121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
      181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      241 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
      301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      781 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      841 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      901 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
      961 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     1021 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     1081 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     1141 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     1201 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     1261 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     1321 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     1381 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     1441 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     1501 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     1561 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     1621 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     1681 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     1741 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     1801 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     1861 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     1921 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     1981 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     2041 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     2101 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     2161 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     2221 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     2281 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     2341 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     2401 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     2461 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     2521 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     2581 aaggctctca agggcatcgg tcgatcgacg ctctccctta tgcgactcct gcattaggaa
     2641 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     2701 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     2761 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     2821 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     2881 gatcgagatc tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt
     2941 ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgtc ccctatacta
     3001 ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa
     3061 gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag
     3121 tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta
     3181 acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt
     3241 ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt
     3301 gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag
     3361 ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt
     3421 gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg
     3481 gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct
     3541 atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc
     3601 tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatctggt tccgcgtcca
     3661 tggtcgaatc aaacaagttt gtacaaaaaa gcaggctcca tgcggccgca gaccctgaag
     3721 ggacaccagg agaagattcg gcagcggcag tccatcctgc ctcctcccca gggcccggcg
     3781 cccatcccct tccagcaccg cggcggggat tccccggagg ccaagaatcg cgtgggcccg
     3841 caggtgccac tcagcgagcc aggtttccgc cgtcgggagt cgcaggagga gccgcgggcc
     3901 gtgctggctc agaagataga gaaggagacg caaatcctca actgcgccct ggacgacatc
     3961 gagtggtttg tggcccggct gcagaaggca gccgaggctt tcaagcagct gaaccagcgg
     4021 aaaaagggga agaagaaggg caagaaggcg ccagcagagg acccagcttt cttgtacaaa
     4081 gtggtttgat tcgacccggg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     4141 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     4201 gggttttttg ctgaaaggag gaactatatc cggatatcca caggacgggt gtggtcgcca
     4261 tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg cggcggccaa
     4321 agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg
     4381 ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc
     4441 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga
     4501 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact
     4561 gtgataaact accgcattaa agcttatcga tgataagctg tcaaacatga gaattcttga
     4621 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4681 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4741 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4801 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4861 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4921 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4981 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5041 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     5101 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5161 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5221 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5281 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5341 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     5401 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5461 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5521 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5581 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5641 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5701 tcatatatac tttagatt
//
