LOCUS       sequence-392526-        6457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GST-Eps8L2_1-496.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6457)
  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 6457)
  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: pGEX-6P-1-Eps8L2_1-496
FEATURES             Location/Qualifiers
     source          1..6457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(181..186,187..202,203..209)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    217..233
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             256..909
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     867..889
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             916..939
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2505..2527)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2672..2690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2758..2862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2863..2931,2932..3723)
                     /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(3081..3100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3894..4482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4383..4402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4636..4653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4726..4803
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4804..5886
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4823..4842)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5935..5940,5941..5958,5959..5965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5973..5989
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5997..6013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5997..6013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6009..6457,1..79)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6029..6046)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6029..6045)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6038..6060)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt
       61 atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
      121 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
      181 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
      241 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      301 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      361 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      421 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      481 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      541 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      601 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      661 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      721 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      781 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      841 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      901 cctccaaaat cggatctgga agttctgttc caggggcccc tgggatccat gagccagtcc
      961 ggggccgtga gctgctgccc gggtgccacc aatggcagcc tgggccggtc cgacggtgtg
     1021 gccaagatga gccccaagga cctgtttgag cagaggaaga agtattccaa ctccaacgtc
     1081 atcatgcacg agacctcgca gtaccacgtc cagcacctgg ccacattcat catggacaag
     1141 agcgaagcca tcacgtctgt ggacgacgcc atccggaagc tggtgcagct gagctccaag
     1201 gagaagatct ggacccagga gatgctgctg caagtcaatg atcagtcgct gcggctgctg
     1261 gacatcgagt cacaggagga gctggaagac ttcccgctgc ccacggtgca gcgcagccag
     1321 acggtcctca accagctgcg ctacccgtct gtgctgctgc tcgtgtgcca ggactcggag
     1381 cagagcaagc cggatgtcca cttcttccac tgcgatgagg tggaggcaga gctggtgcac
     1441 gaggacatcg agagcgcgtt ggccgactgc cggctgggca agaagatgcg gccgcagacc
     1501 ctgaagggac accaggagaa gattcggcag cggcagtcca tcctgcctcc tccccagggc
     1561 ccggcgccca tccccttcca gcaccgcggc ggggattccc cggaggccaa gaatcgcgtg
     1621 ggcccgcagg tgccactcag cgagccaggt ttccgccgtc gggagtcgca ggaggagccg
     1681 cgggccgtgc tggctcagaa gatagagaag gagacgcaaa ttctcaactg cgccctggac
     1741 gacatcgagt ggtttgtggc ccggctgcag aaggcagccg aggctttcaa gcagctgaac
     1801 cagcggaaaa aggggaagaa gaagggcaag aaggcgccag cagagggcgt cctcacactg
     1861 cgggcacggc ccccctctga gggcgagttc atcgactgct tccagaaaat caagctggcg
     1921 attaacttgc tggcaaagct gcagaagcac atccagaacc ccagcgccgc ggagctcgtg
     1981 cacttcctct tcgggcctct ggacctgatc gtcaacacct gcagtggccc agacatcgca
     2041 cgctccgtct cctgcccact gctctcccga gatgccgtgg acttcctgcg cggccacctg
     2101 gtccctaagg agatgtcgct gtgggagtca ctgggagaga gctggatgcg gccccgttcc
     2161 gagtggccgc gggagccaca ggtgcccctc tacgtgccca agttccacag cggctgggag
     2221 cctcctgtgg atgtgctgca ggaggccccc tgggaggtgg aggggctggc gtctgccccc
     2281 atcgaggagg tgagtccagt gtcgaggcaa agtatccgga actcccagaa gcacagcccc
     2341 acttcagagc ccaccccccc gggggatgcc ctaccaccag tcagctcccc acatactcac
     2401 aggggctacc agccaacacc agccatggcc aaatattaac tcgagcggcc gcatcgtgac
     2461 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2521 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2581 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     2641 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2701 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2881 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2941 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3001 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3061 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3121 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3181 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3241 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3301 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3361 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3421 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3481 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3541 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3601 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3661 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3721 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3781 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3841 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3901 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3961 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4021 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4081 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4141 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4201 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4261 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4321 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4381 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4441 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4501 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4561 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4621 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4681 tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg
     4741 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     4801 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     4861 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     4921 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     4981 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     5041 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     5101 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     5161 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     5221 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     5281 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     5341 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     5401 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     5461 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     5521 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     5581 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     5641 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     5701 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     5761 cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca atacgcaaac cgcctctccc
     5821 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5881 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca
     5941 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6001 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
     6061 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     6121 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     6181 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     6241 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     6301 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
     6361 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     6421 gccagacgcg aattattttt gatggcgttg gaattag
//
