LOCUS       sequence-490278-        8973 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  Expression of wild-type slingshot 3, a cofilin pSer3 phosphatase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8973)
  AUTHORS   Ohta Y, Kousaka K, Nagata-Ohashi K, Ohashi K, Muramoto A, Shima Y, 
            Niwa R, Uemura T, Mizuno K
  TITLE     Differential activities, subcellular distribution and tissue 
            expression patterns of three members of Slingshot family 
            phosphatases that dephosphorylate cofilin.
  JOURNAL   Genes Cells. 2003 Oct;8(10):811-24. doi: 
            10.1046/j.1365-2443.2003.00678.x.
  PUBMED    14531860
REFERENCE   2  (bases 1 to 8973)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1046/j.1365-2443.2003.00678"; journalName: "Genes Cells"; date: 
            "2003-10"; volume: "8"; issue: "10"; pages: "811-24"
COMMENT     Sequence Label: SSH-3 WT
FEATURES             Location/Qualifiers
     source          1..8973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(22..402)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(84..105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     296..315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   440..542
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     630..780
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        838..1141
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1142..1345
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1296..1316
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2857..2886
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2902..2919
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3059..3180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3096..3115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3150..3169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6371..6473
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(6562..6579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6733..7321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6813..6832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             8153..8947
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             8153..8947
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8207..8226)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8817..8836
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
       61 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      121 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      181 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      241 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      301 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      361 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggat
      421 cgaattaatt cttaattaac atcatcaata atatacctta ttttggattg aagccaatat
      481 gataatgagg gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt
      541 agtagtgtgg cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt
      601 ggcaaaagtg acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg
      661 ttttaggcgg atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg
      721 ggaaaactga ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat
      781 actgtaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      841 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      901 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      961 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     1021 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1081 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1141 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1201 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1261 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1321 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agagatctgg
     1381 taccgtcgac gcggccgctc gagcctaagc ttgatggccc tggtcacagt gagccgttcg
     1441 cccccgggca gcggcgcctc cacgcccgtg gggccctggg accaggcggt ccagcgaagg
     1501 agtcgactcc agcgaaggca gagctttgcg gtgctccgtg gggctgtcct gggactgcag
     1561 gatggagggg acaatgatga tgcagtagag gccagttctg agccaacaga gaaggccccg
     1621 agtgaggagg agctccacgg ggaccagaca gacttcgggc aaggatccca gagtccccag
     1681 aagcaggagg agcagaggca gcacctgcac ctcatggtac agctgctgag gccgcaggat
     1741 gacatccgcc tggcagccca gctggaggca ccccggcctc cccggctccg ctacctgctg
     1801 gtagtttcta cacgagaagg agaaggtctg agccaggatg agacggtcct cctgggcgtg
     1861 gatttccctg acagcagctc ccccagctgc accctgggcc tggtcttgcc cctctggagt
     1921 gacacccagg tgtacttaga tggagacggg ggcttcagcg tgacgtctgg tgggcaaagc
     1981 cggatcttca agcccatctc catccagacc atgtgggcca cactccaggt attgcaccaa
     2041 gcatgtgagg cagctctagg cagcggcctt gtaccgggtg gcagtgccct cacctgggcc
     2101 agccactacc aggagagact gaactccgaa cagagctgcc tcaatgagtg gacggctatg
     2161 gccgacctgg agtctctgcg gcctcccagc gccgagcctg gcgggtcctc agaacaggag
     2221 cagatggagc aggcgatccg tgctgagctg tggaaagtgt tggatgtcag tgacctggag
     2281 agtgtcactt ccaaagagat ccgccaggct ctggagctgc gcctggggct ccccctccag
     2341 cagtaccgtg acttcatcga caaccagatg ctgctgctgg tggcacagcg ggaccgagcc
     2401 tcccgcatct tcccccacct ctacctgggc tcagagtgga acgcagcaaa cctggaggag
     2461 ctgcagagga acagggtcac ccacatcttg aacatggccc gggagattga caacttctac
     2521 cctgagcgct tcacctacca caatgtgcgc ctctgggatg aggagtcggc ccagctgctg
     2581 ccgcactgga aggagacgca ccgcttcatt gaggctgcaa gagcacaggg cacccacgtg
     2641 ctggtccact gcaagatggg cgtcagccgc tcagcggcca cagtgctggc ctatgccatg
     2701 aagcagtacg aatgcagcct ggagcaggcc ctgcgccacg tgcaggagct ccggcccatc
     2761 gcccgcccca accctggctt cctgcgccag ctgcagatct accagggcat cctgacggcc
     2821 agaacctcgc ggccgctcga gtctagaggg cccttcgaac aaaaactcat ctcagaagag
     2881 gatctgaata tgcataccgg tcatcatcac catcaccatt gagtttatcc gatccaccgg
     2941 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3001 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3061 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3121 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3181 acgcggatct gggcgtggtt aagggtggga aagaatatat aaggtggggg tcttatgtag
     3241 ttttgtatct gttttgcagc agccgccgcc gccatgagca ccaactcgtt tgatggaagc
     3301 attgtgagct catatttgac aacgcgcatg cccccatggg ccggggtgcg tcagaatgtg
     3361 atgggctcca gcattgatgg tcgccccgtc ctgcccgcaa actctactac cttgacctac
     3421 gagaccgtgt ctggaacgcc gttggagact gcagcctccg ccgccgcttc agccgctgca
     3481 gccaccgccc gcgggattgt gactgacttt gctttcctga gcccgcttgc aagcagtgca
     3541 gcttcccgtt catccgcccg cgatgacaag ttgacggctc ttttggcaca attggattct
     3601 ttgacccggg aacttaatgt cgtttctcag cagctgttgg atctgcgcca gcaggtttct
     3661 gccctgaagg cttcctcccc tcccaatgcg gtttaaaaca taaataaaaa accagactct
     3721 gtttggattt ggatcaagca agtgtcttgc tgtctttatt taggggtttt gcgcgcgcgg
     3781 taggcccggg accagcggtc tcggtcgttg agggtcctgt gtattttttc caggacgtgg
     3841 taaaggtgac tctggatgtt cagatacatg ggcataagcc cgtctctggg gtggaggtag
     3901 caccactgca gagcttcatg ctgcggggtg gtgttgtaga tgatccagtc gtagcaggag
     3961 cgctgggcgt ggtgcctaaa aatgtctttc agtagcaagc tgattgccag gggcaggccc
     4021 ttggtgtaag tgtttacaaa gcggttaagc tgggatgggt gcatacgtgg ggatatgaga
     4081 tgcatcttgg actgtatttt taggttggct atgttcccag ccatatccct ccggggattc
     4141 atgttgtgca gaaccaccag cacagtgtat ccggtgcact tgggaaattt gtcatgtagc
     4201 ttagaaggaa atgcgtggaa gaacttggag acgcccttgt gacctccaag attttccatg
     4261 cattcgtcca taatgatggc aatgggccca cgggcggcgg cctgggcgaa gatatttctg
     4321 ggatcactaa cgtcatagtt gtgttccagg atgagatcgt cataggccat ttttacaaag
     4381 cgcgggcgga gggtgccaga ctgcggtata atggttccat ccggcccagg ggcgtagtta
     4441 ccctcacaga tttgcatttc ccacgctttg agttcagatg gggggatcat gtctacctgc
     4501 ggggcgatga agaaaacggt ttccggggta ggggagatca gctgggaaga aagcaggttc
     4561 ctgagcagct gcgacttacc gcagccggtg ggcccgtaaa tcacacctat taccggctgc
     4621 aactggtagt taagagagct gcagctgccg tcatccctga gcaggggggc cacttcgtta
     4681 agcatgtccc tgactcgcat gttttccctg accaaatccg ccagaaggcg ctcgccgccc
     4741 agcgatagca gttcttgcaa ggaagcaaag tttttcaacg gtttgagacc gtccgccgta
     4801 ggcatgcttt tgagcgtttg accaagcagt tccaggcggt cccacagctc ggtcacctgc
     4861 tctacggcat ctcgatccag catatctcct cgtttcgcgg gttggggcgg ctttcgctgt
     4921 acggcagtag tcggtgctcg tccagacggg ccagggtcat gtctttccac gggcgcaggg
     4981 tcctcgtcag cgtagtctgg gtcacggtga aggggtgcgc tccgggctgc gcgctggcca
     5041 gggtgcgctt gaggctggtc ctgctggtgc tgaagcgctg ccggtcttcg ccctgcgcgt
     5101 cggccaggta gcatttgacc atggtgtcat agtccagccc ctccgcggcg tggcccttgg
     5161 cgcgcagctt gcccttggag gaggcgccgc acgaggggca gtgcagactt ttgagggcgt
     5221 agagcttggg cgcgagaaat accgattccg gggagtaggc atccgcgccg caggccccgc
     5281 agacggtctc gcattccacg agccaggtga gctctggccg ttcggggtca aaaaccaggt
     5341 ttcccccatg ctttttgatg cgtttcttac ctctggtttc catgagccgg tgtccacgct
     5401 cggtgacgaa aaggctgtcc gtgtccccgt atacagactt gagagggagt ttaaacgaat
     5461 tcaatagctt gttgcatggg cggcgatata aaatgcaagg tgctgctcaa aaaatcaggc
     5521 aaagcctcgc gcaaaaaaga aagcacatcg tagtcatgct catgcagata aaggcaggta
     5581 agctccggaa ccaccacaga aaaagacacc atttttctct caaacatgtc tgcgggtttc
     5641 tgcataaaca caaaataaaa taacaaaaaa acatttaaac attagaagcc tgtcttacaa
     5701 caggaaaaac aacccttata agcataagac ggactacggc catgccggcg tgaccgtaaa
     5761 aaaactggtc accgtgatta aaaagcacca ccgacagctc ctcggtcatg tccggagtca
     5821 taatgtaaga ctcggtaaac acatcaggtt gattcacatc ggtcagtgct aaaaagcgac
     5881 cgaaatagcc cgggggaata catacccgca ggcgtagaga caacattaca gcccccatag
     5941 gaggtataac aaaattaata ggagagaaaa acacataaac acctgaaaaa ccctcctgcc
     6001 taggcaaaat agcaccctcc cgctccagaa caacatacag cgcttccaca gcggcagcca
     6061 taacagtcag ccttaccagt aaaaaagaaa acctattaaa aaaacaccac tcgacacggc
     6121 accagctcaa tcagtcacag tgtaaaaaag ggccaagtgc agagcgagta tatataggac
     6181 taaaaaatga cgtaacggtt aaagtccaca aaaaacaccc agaaaaccgc acgcgaacct
     6241 acgcccagaa acgaaagcca aaaaacccac aacttcctca aatcgtcact tccgttttcc
     6301 cacgttacgt cacttcccat tttaagaaaa ctacaattcc caacacatac aagttactcc
     6361 gccctaaaac ctacgtcacc cgccccgttc ccacgccccg cgccacgtca caaactccac
     6421 cccctcatta tcatattggc ttcaatccaa aataaggtat attattgatg atgttaatta
     6481 acatgcatgg atccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6541 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7141 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7801 cattgctgca gccatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa
     7861 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     7921 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     7981 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     8041 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     8101 atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     8161 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     8221 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     8281 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     8341 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     8401 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     8461 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     8521 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     8581 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     8641 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     8701 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     8761 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     8821 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     8881 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     8941 cttctgaatt ttgttaaaat ttttgttaaa tca
//
