LOCUS       sequence-386579-        7499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2-FL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7499)
  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 7499)
  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
FEATURES             Location/Qualifiers
     source          1..7499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      134..722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     623..642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     876..893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    908..1047
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(973..992)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1133..1155
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1149..1340)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2802..2821
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2905..2924
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2905..2923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2955..2977
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2985..3638
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    3672..3696
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5841..5865)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5929..5947)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5943..5990
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6356..6361,6362..6378,6379..6384))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6411..6429)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6497..6601
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6602..6670,6671..7462)
                     /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(6820..6839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
       61 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
      121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      241 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
      301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      781 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      841 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      901 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
      961 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     1021 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     1081 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     1141 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     1201 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     1261 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     1321 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     1381 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     1441 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     1501 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     1561 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     1621 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     1681 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     1741 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc
     1801 acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct ggagatggcg
     1861 gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct ccgcaagaat
     1921 tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg gcttccattc
     1981 aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag acaaggtata
     2041 gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg gcataaatcg
     2101 ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg agcgatcctt
     2161 gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc aacgcgggca
     2221 tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag cctcgcgtcg
     2281 cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata atggcctgct
     2341 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     2401 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     2461 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     2521 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     2581 aggctctcaa gggcatcggt cgatcgacgc tctcccttat gcgactcctg cattaggaag
     2641 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     2701 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     2761 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     2821 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     2881 atcgagatct cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
     2941 cctctagaaa taattttgtt taactttaag aaggagatat acatatgtcc cctatactag
     3001 gttattggaa aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag
     3061 aaaaatatga agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt
     3121 ttgaattggg tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa
     3181 cacagtctat ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc
     3241 caaaagagcg tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg
     3301 tttcgagaat tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc
     3361 tacctgaaat gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg
     3421 atcatgtaac ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg
     3481 acccaatgtg cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta
     3541 tcccacaaat tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct
     3601 ggcaagccac gtttggtggt ggcgaccatc ctccaaaatc ggatctggtt ccgcgtccat
     3661 ggtcgaatca aacaagtttg tacaaaaaag caggcttgag ccagtccggg gccgtgagct
     3721 gctgcccggg tgccaccaat ggcagcctgg gccggtccga cggtgtggcc aagatgagcc
     3781 ccaaggacct gtttgagcag aggaagaagt attccaactc caacgtcatc atgcacgaga
     3841 cctcgcagta ccacgtccag cacctggcca cattcatcat ggacaagagc gaagccatca
     3901 cgtctgtgga cgacgccatc cggaagctgg tgcagctgag ctccaaggag aagatctgga
     3961 cccaggagat gctgctgcag gtgaacgacc agtcgctgcg gctgctggac atcgagtcac
     4021 aggaggagct ggaagacttc ccgctgccca cggtgcagcg cagccagacg gtcctcaacc
     4081 agctgcgcta cccgtctgtg ctgctgctcg tgtgccagga ctcggagcag agcaagccgg
     4141 atgtccactt cttccactgc gatgaggtgg aggcagagct ggtgcacgag gacatcgaga
     4201 gcgcgttggc cgactgccgg ctgggcaaga agatgcggcc gcagaccctg aagggacacc
     4261 aggagaagat tcggcagcgg cagtccatcc tgcctcctcc ccagggcccg gcgcccatcc
     4321 ccttccagca ccgcggcggg gattccccgg aggccaagaa tcgcgtgggc ccgcaggtgc
     4381 cactcagcga gccaggtttc cgccgtcggg agtcgcagga ggagccgcgg gccgtgctgg
     4441 ctcagaagat agagaaggag acgcaaatcc tcaactgcgc cctggacgac atcgagtggt
     4501 ttgtggcccg gctgcagaag gcagccgagg ctttcaagca gctgaaccag cggaaaaagg
     4561 ggaagaagaa gggcaagaag gcgccagcag agggcgtcct cacactgcgg gcacggcccc
     4621 cctctgaggg cgagttcatc gactgcttcc agaaaatcaa gctggcgatt aacttgctgg
     4681 caaagctgca gaagcacatc cagaacccca gcgccgcgga gctcgtgcac ttcctcttcg
     4741 ggcctctgga cctgatcgtc aacacctgca gtggcccaga catcgcacgc tccgtctcct
     4801 gcccactgct ctcccgagat gccgtggact tcctgcgcgg ccacctggtc cctaaggaga
     4861 tgtcgctgtg ggagtcactg ggagagagct ggatgcggcc ccgttccgag tggccgcggg
     4921 agccacaggt gcccctctac gtgcccaagt tccacagcgg ctgggagcct cctgtggatg
     4981 tgctgcagga ggccccctgg gaggtggagg ggctggcgtc tgcccccatc gaggaggtga
     5041 gtccagtgtc gaggcaaagt atccggaact cccagaagca cagccccact tcagagccca
     5101 cccccccggg ggatgcccta ccaccagtca gctccccaca tactcacagg ggctaccagc
     5161 caacaccagc catggccaaa tatgtgaaaa ttctctatga cttcacagcc cgaaatgcca
     5221 acgagctatc ggtgctcaag gatgaggtcc tagaggtgct ggaggacggc cggcagtggt
     5281 ggaagctgcg cagccgcagc ggccaggcgg ggtacgtgcc ctgcaacatc ctaggcgagg
     5341 cgcgaccgga ggacgccggc gccccgttcg agcaggccgg tcagaagtac tggggccccg
     5401 ccagcccgac ccacaagcta cccccaagct tcccgggcaa taaggacgaa ctcatgcagc
     5461 acatggacga ggtcaacgac gagctcatcc gcaagatatc gaatataagg gcgcagccac
     5521 agaggcactt ccgcgtggag cgcagccagc ccgtgagcca gccgctcacc tacgagtcgg
     5581 gtccggacga ggtccgcgcc tggctggaag ccaaggcctt cagcccgcgg atcgtggaga
     5641 acctgggcat cctgaccggg ccgcagctct tctccctcaa caaggaggag ctgaagaaag
     5701 tgtgcggcga ggagggcgtc cgcgtgtaca gccagctcac catgcagaag gccttcctgg
     5761 agaagcagca aagtgggtcg gagctggaag aactcatgaa caagtttcat tccatgaatc
     5821 agaggagggg ggaggacagc acccagcttt cttgtacaaa gtggtttgat tcgacccggg
     5881 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     5941 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     6001 gaactatatc cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg
     6061 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
     6121 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
     6181 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
     6241 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
     6301 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
     6361 agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat
     6421 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6481 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6541 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6601 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6661 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6721 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6781 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6841 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6901 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6961 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7021 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     7081 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7141 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7201 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7261 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7321 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7381 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7441 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattg
//
