LOCUS       sequence-368873-        8820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Plekhh1 with 5 silent mutations labelled with 
            Cherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8820)
  AUTHORS   Prosperi MT, Pernier J, Lachuer H, Coudrier E
  TITLE     Plekhh1, a partner of myosin 1 and an effector of EphB2, controls 
            the cortical actin network during cell repulsion.
  JOURNAL   J Cell Sci. 2021 Dec 1;134(23). pii: 272686. doi: 
            10.1242/jcs.258802. Epub 2021 Dec 6.
  PUBMED    34723325
REFERENCE   2  (bases 1 to 8820)
  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"; journalName: "J Cell 
            Sci."; date: "2021-12-1"; volume: "134(23). pii: 272686. doi"; 
            pages: " 10.1242/jcs.258802. Epub 2021 Dec 6"
COMMENT     Sequence Label: Cherry-Plekhh1 5 silent mutations
FEATURES             Location/Qualifiers
     source          1..8820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      613..622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             619..1326
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(764..782)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1032..1051
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5608..5729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5645..5664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5699..5718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5736..6191)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5873..5894)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6085..6104
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6218..6322
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6324..6681
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6324..6515
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6347..6367)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6532..6667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6716..7510
                     /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             6716..7510
                     /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(6770..6789)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7380..7399
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7698..7717)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7742..7789
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8118..8706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8607..8626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgttt
      601 aaacttaagc ttgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
      661 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      721 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
      781 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
      841 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
      901 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
      961 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     1021 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     1081 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     1141 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     1201 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     1261 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     1321 tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctatggcaga actcaaggtg
     1381 gaggcgccgg ccagcgtaga ctggcagaaa cgctgcctga ccctggaaac tcagcttttc
     1441 cggttccgcc tacaggccag caagataagg gagctgctgg ctgacaagat gcaggagctg
     1501 gagcagaggc tgctggaggc agagcagaga gcagagaacg ctgagaccca ggtgggtgtc
     1561 atggaagaga aggtaaaact atccaatctg aagaatgtgg actctgaggg gagcctgcac
     1621 cggaaatacc aagaattgct gaaagccata aagggcaaag atgagctcat cagccagcta
     1681 gaggctcagc tggagaagca gaagcagatg agggcagagg aagcaaaaac tgttcaagaa
     1741 aaagctgcaa agatcaagga atgggtgaca ctcaagttgg caaagcttga gatggagaat
     1801 cagcatctga aaagccataa tcagcgcctg gtggagcagg tgggatccct tcaagatgcg
     1861 ctagaagcta ttcagatagc tccttcacgg aagctgctgg tgccccccta cggagctgca
     1921 gagcaggatt ctgtcccttc agagccggga atccagccta tgggccagga cagtggctcc
     1981 caggcccagg gtctgaaggc agctgtgctt gcaccttccc caggtgccct gcagagcaag
     2041 gactctgttt ctgaagcagc aagccccttg gaggattcta gttccagcac ggtccattct
     2101 ggggaaacag tagaggccaa gccccttcaa cctcatctgg gaagagagag ccctccccac
     2161 cagccatgca tgaagcttct taccttcaga tgtagttcag cttcctgggg tgagggtctg
     2221 gttactgctc agagagggat gctccctggg acaaagacct ctgccaggga aggtggtcct
     2281 ggcagcagtc tgaccctacc aaaggtgcgg gctcctggca ccccgcggga cagcatccag
     2341 ttggccaaaa ggcaccacag ccagccccag gtgggccatg ggcactttgg ccgtgtggta
     2401 aacattgaga ctgaggcctt ctcagccctc cacccctctg gccttcctga gctggagtcc
     2461 cgagctaggt cccgggagga accagagaag atggagatgg aggagccacc cccagcaggg
     2521 aagaatgagg aaagagagag cccaaaggcc cttggagctg agctggagga agtggagctg
     2581 ggtaacaagc cacctacacc cccgctgcac cagttttcat cctgggagag ccggatctat
     2641 gctgtggcca catcgggcat gcggctctca gatatgtctc ccagaagtaa tactgcatgc
     2701 tgcgcttcaa gccctcctgc ccttgtttcc cctgggtctt tctctggcct ggtctacaag
     2761 aatgtcactg tgcctgtcta cacagcactg aaggggagag ctacacagat cagcaacatg
     2821 ccctttatgg acgagtcctc tgggtctgac gatgactgca gctctcaggc gagtttccga
     2881 atctcggtcc cctcctctga gtccagaaaa acaagtggtc taggcagccc ccgggccatc
     2941 aagagaggcg tctccatgtc ctcactgagc tccgagggtg actacgccat ccccccggac
     3001 gcctgctcac tggacagtga ctactcagag cctgagcaca aactgcagcg cacctcatcc
     3061 tactccaccg acgggctggg cctgggcggg gagtcactgg agaagtcggg ctacctgctg
     3121 aaaatgggga gccaggtgaa gacgtggaag aggcgctggt ttgtcctgag acagggacag
     3181 attatgtact acaagtcccc gagtgatgtc atccggaaac ctcaaggcca agtggatctg
     3241 aactcccgct gccaaattgt tcgaggggag ggttcacaga cgtttcagct catctctgag
     3301 aagaaaacct actacctgac ggccgattca cccagcctgc tggaggagtg gatccgagta
     3361 ctccagagcc tgctgaaggt gcaggccacc gggcctccag ctctgcttcg gggtggcacc
     3421 aagcccaccg tgaagggctg gctgaccaag gtaaagcatg gccactccaa ggtggtctgg
     3481 tgcgctcttg ttgggaaaat cttctactac tatcggagcc atgaggacaa gcgacccctg
     3541 ggctgcctgc ctgtgcggga tgcgcacata gaggaagtag atcgatcctg tgactcagac
     3601 gaggactatg aggctggagg aaccagacgg ttgctttcct cccactgcac cctggtgatc
     3661 caccccacag agcacagccc cacctacctc ctcattggca ccaagcatga aaaggatacg
     3721 tggctctacc acctcacagt ggctgcaggt ggcagcagtg ccaaggtggg cactgcctat
     3781 gagcagctca ttggaaaact gatggatggt gaaggagatc cagattcgcc gctctggagg
     3841 caccccatgc tgtgctacag caaagacggc ctatacgcct ccctcaccac cctgccctct
     3901 gaggccttgc agacggaggc cctcaagctc ttcaagtcct gccagctctt catcaacgtg
     3961 ccggtggaag ctgcctcggt ggactaccat gtgtccctgg cccagaccgc actgcaggtc
     4021 tgcctggttc accccgagct gcagagtgag atctactgcc aactcatgaa gcagaccagc
     4081 tgccgcccac ctcagaagta ctccctcatg cagtgctggc agctcctcgc tctgtgtgct
     4141 ccacttttcc tgcctcagca tcacttcctc tggtatgtca agcagcagct ccaacgccat
     4201 gcagatccca gaagtgaaac tggccagtat gccacctact gccagcgggc agtggagcgg
     4261 accctgcgga ccggggagcg ggaagccagg ccatcgcgca tggaagtggt gtccatcctg
     4321 ctgcgtaacc ccttccacca ctccttgccc ttcagcatcc ccgtgcactt taccaacggg
     4381 acttaccatg tggttggttt tgacggctct tccacggttg atgagttcct ccagcggctg
     4441 aaccaggaga taggcatgag aaagccatcc cactctggct ttgccctctt cacggacgat
     4501 ccctcgggca gggacctgga gcactgcctg cagggaagtg tcaagatctg tgatgccatc
     4561 tccaagtggg aacaagccat gaaggagctg caccccggaa agtctgaggg tgggacacgc
     4621 gtcgtgaagc tgatgtacaa gaacaggctg tactttcgca gtcaagtcaa aggggagacg
     4681 gaccgagaac ggctgctcct tgcctctcaa accagtagag agatagtggc agggaggttt
     4741 cctatcaaca aggaattggc tcttgagatg gctgccctga tggcccaggt agaatatggg
     4801 gacttggaga agcctgccct gccaggccct ggaggcacat cccctgccaa ggctcagcat
     4861 cttctccagc aggtcctaga caggttccac cccaggcgct atagacatgg ggcccccgct
     4921 gaacagctga ggcacctggc agatatgttg accacaaaat gggcaacatt gcaaggatgc
     4981 tcccctcctg agtgcatccg catctacctg accgtggcca ggaaatggcc tttctttggt
     5041 gctaaacttt ttgctgctca gcctgcccag ctgtcttcca aggagaacgc tctggtgtgg
     5101 attgctgtga atgaggatgg cgtcagcatc ctggaccaca acactatgca agtgcacatc
     5161 acttacccct actcttcagt gacaacgttt ggtggctgca gggatgactt catgcttgtg
     5221 attagatcta tcccagacaa gagctctgga aaaagccaca ttgagaagtt gatcttccgg
     5281 atggctgctc ccaagattgc agaagccacc ttcatcatgg ccagctatat gaaccattgc
     5341 actacaactg tgaacccccc caccaaccca cccggagcct gccagctgtg ggaactggat
     5401 ggacgacagt tcttttcttc tgtttcctgt gctaccaagg ggccaacgtt gctgtgagtc
     5461 gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc
     5521 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     5581 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5641 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5701 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     5761 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5821 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5881 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5941 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6001 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6061 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6121 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6181 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6241 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6301 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     6361 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6421 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6481 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6541 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6601 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6661 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     6721 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     6781 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     6841 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     6901 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     6961 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     7021 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     7081 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     7141 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     7201 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     7261 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7321 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7381 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7441 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     7501 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     7561 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     7621 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7681 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     7741 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     7801 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     7861 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     7921 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     7981 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8041 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8101 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8161 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8221 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8281 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8341 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8401 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8461 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8521 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8581 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8641 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8701 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8761 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
