LOCUS       sequence-385270-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_367-496.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  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 5751)
  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_367-496
FEATURES             Location/Qualifiers
     source          1..5751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(2..590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(82..101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(761..1552,1553..1621))
                     /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     1384..1403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1622..1726)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1794..1812
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1839..1844,1845..1861,1862..1867)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2233..2280)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2276..2294
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    2358..2382
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2779..2803)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2837..3490)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     RBS             3498..3520
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3551..3570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3552..3570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3654..3673)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5135..5326
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5320..5342)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5428..5567
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5483..5502
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5582..5599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
       61 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      181 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      421 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
      481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
      961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     1081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     1381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1741 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1801 gtatcacgag gccctttcgt cttcaagaat tctcatgttt gacagcttat catcgataag
     1861 ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa
     1921 tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     1981 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     2041 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     2101 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     2161 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatatc cggatatagt
     2221 tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag
     2281 ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg
     2341 atcccgggtc gaatcaaacc actttgtaca agaaagctgg gtcatatttg gccatggctg
     2401 gtgttggctg gtagcccctg tgagtatgtg gggagctgac tggtggtagg gcatcccccg
     2461 ggggggtggg ctctgaagtg gggctgtgct tctgggagtt ccggatactt tgcctcgaca
     2521 ctggactcac ctcctcgatg ggggcagacg ccagcccctc cacctcccag ggggcctcct
     2581 gcagcacatc cacaggaggc tcccagccgc tgtggaactt gggcacgtag aggggcacct
     2641 gtggctcccg cggccactcg gaacggggcc gcatccagct ctctcccagt gactcccaca
     2701 gcgacatctc cttagggacc aggtggccgc gcaggaagtc cacggcatct cgggagagca
     2761 gtgggcagga gaccatggag cctgcttttt tgtacaaact tgtttgattc gaccatggac
     2821 gcggaaccag atccgatttt ggaggatggt cgccaccacc aaacgtggct tgccagccct
     2881 gcaaaggcca tgctatatac ttgctggatt tcaagtactt atcaatttgt gggatagctt
     2941 caatacgttt tttaaaacaa actaattttg ggaacgcatc caggcacatt gggtccatgt
     3001 ataaaacaac atcaagagcg tcatacaaca tgaagtcagg atgggttaca tgatcaccat
     3061 ttaaatatgt tttatgacat aaacgatctt cgaacatttt cagcatttca ggtagcttgc
     3121 taagaaaatc aactttgaga gtttcaaagt ctttactata tgcaattctc gaaacaccgt
     3181 atctaatatc caaaaccgct ccttcaagca ttgaaatctc tgcacgctct tttggacaac
     3241 cacccaacat gttgtgcttg tcagctatat aacgtatgat ggccatagac tgtgttaatt
     3301 taacatcacc atcaatataa taaggaagat tgggaaactc caaacccaat tcaaactttt
     3361 tgtttcgcca tttatcacct tcatcgcgct catacaaatg ctcttcatat ttttcttcaa
     3421 gatattccaa aagaagtcga gtgggttgca caaggccctt aattttccaa taacctagta
     3481 taggggacat atgtatatct ccttcttaaa gttaaacaaa attatttcta gagggaaacc
     3541 gttgtggtct ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta
     3601 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     3661 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     3721 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     3781 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     3841 aatgcaggag tcgcataagg gagagcgtcg atcgaccgat gcccttgaga gccttcaacc
     3901 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     3961 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     4021 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     4081 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     4141 ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc
     4201 gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg
     4261 cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat
     4321 cgctcgcggc tcttaccagc ctaacttcga tcattggacc gctgatcgtc acggcgattt
     4381 atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc
     4441 ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg
     4501 aagccggcgg cacctcgcta acggattcac cactccaaga attggagcca atcaattctt
     4561 gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat
     4621 ctccagcagc cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat
     4681 gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca
     4741 gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac
     4801 ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa
     4861 gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg
     4921 tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg
     4981 gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg
     5041 ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc
     5101 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
     5161 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
     5221 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
     5281 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5341 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     5401 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     5461 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc
     5521 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     5581 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5641 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5701 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg t
//
