LOCUS       sequence-368847-        8823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Plekhh1 labelled with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8823)
  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 8823)
  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: EGFP-Plekhh1
FEATURES             Location/Qualifiers
     source          1..8823
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5611..5732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5648..5667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5702..5721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5739..6194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5876..5897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6088..6107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6221..6325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6327..6684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6327..6518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6350..6370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6535..6670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6719..7513
                     /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             6719..7513
                     /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(6773..6792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7383..7402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7701..7720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7745..7792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8121..8709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8610..8629
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctatggc agaactcaag
     1381 gtggaggcgc cggccagcgt agactggcag aaacgctgcc tgaccctgga aactcagctt
     1441 ttccggttcc gcctacaggc cagcaagata agggagctgc tggctgacaa gatgcaggag
     1501 ctggagcaga ggctgctgga ggcagagcag agagcagaga acgctgagac ccaggtgggt
     1561 gtcatggaag agaaggtaaa actatccaat ctgaagaatg tggactctga ggggagcctg
     1621 caccggaaat accaagaatt gctgaaagcc ataaagggca aagatgagct catcagccag
     1681 ctagaggctc agctggagaa gcagaagcag atgagggcag aggaagcaaa aactgttcaa
     1741 gaaaaagctg caaagatcaa ggaatgggtg acactcaagt tggcaaagct tgagatggag
     1801 aatcagcatc tgaaaagcca taatcagcgc ctggtggagc aggtgggatc ccttcaagat
     1861 gcgctagaag ctattcagat agctccttca cggaagctgc tggtgccccc ctacggagct
     1921 gcagagcagg attctgtccc ttcagagccg ggaatccagc ctatgggcca ggacagtggc
     1981 tcccaggccc agggtctgaa ggcagctgtg cttgcacctt ccccaggtgc cctgcagagc
     2041 aaggactctg tttctgaagc agcaagcccc ttggaggatt ctagttccag cacggtccat
     2101 tctggggaaa cagtagaggc caagcccctt caacctcatc tgggaagaga gagccctccc
     2161 caccagccat gcatgaagct tcttaccttc agatgtagtt cagcttcctg gggtgagggt
     2221 ctggttactg ctcagagagg gatgctccct gggacaaaga cctctgccag ggaaggtggt
     2281 cctggcagca gtctgaccct accaaaggtg cgggctcctg gcaccccgcg ggacagcatc
     2341 cagttggcca aaaggcacca cagccagccc caggtgggcc atgggcactt tggccgtgtg
     2401 gtaaacattg agactgaggc cttctcagcc ctccacccct ctggccttcc tgagctggag
     2461 tcccgagcta ggtcccggga ggaaccagag aagatggaga tggaggagcc acccccagca
     2521 gggaagaatg aggaaagaga gagcccaaag gcccttggag ctgagctgga ggaagtggag
     2581 ctgggtaaca agccacctac acccccgctg caccagtttt catcctggga gagccggatc
     2641 tatgctgtgg ccacatcggg catgcggctc tcagatatgt ctcccagaag taatactgca
     2701 tgctgcgctt caagccctcc tgcccttgtt tcccctgggt ctttctctgg cctggtctac
     2761 aagaatgtca ctgtgcctgt ctacacagca ctgaagggga gagctacaca gatcagcaac
     2821 atgcccttta tggacgagtc ctctgggtct gacgatgact gcagctctca ggcgagtttc
     2881 cgaatctcgg tcccctcctc tgagtccagg aagaccagcg gactaggcag cccccgggcc
     2941 atcaagagag gcgtctccat gtcctcactg agctccgagg gtgactacgc catccccccg
     3001 gacgcctgct cactggacag tgactactca gagcctgagc acaaactgca gcgcacctca
     3061 tcctactcca ccgacgggct gggcctgggc ggggagtcac tggagaagtc gggctacctg
     3121 ctgaaaatgg ggagccaggt gaagacgtgg aagaggcgct ggtttgtcct gagacaggga
     3181 cagattatgt actacaagtc cccgagtgat gtcatccgga aacctcaagg ccaagtggat
     3241 ctgaactccc gctgccaaat tgttcgaggg gagggttcac agacgtttca gctcatctct
     3301 gagaagaaaa cctactacct gacggccgat tcacccagcc tgctggagga gtggatccga
     3361 gtactccaga gcctgctgaa ggtgcaggcc accgggcctc cagctctgct tcggggtggc
     3421 accaagccca ccgtgaaggg ctggctgacc aaggtaaagc atggccactc caaggtggtc
     3481 tggtgcgctc ttgttgggaa aatcttctac tactatcgga gccatgagga caagcgaccc
     3541 ctgggctgcc tgcctgtgcg ggatgcgcac atagaggaag tagatcgatc ctgtgactca
     3601 gacgaggact atgaggctgg aggaaccaga cggttgcttt cctcccactg caccctggtg
     3661 atccacccca cagagcacag ccccacctac ctcctcattg gcaccaagca tgaaaaggat
     3721 acgtggctct accacctcac agtggctgca ggtggcagca gtgccaaggt gggcactgcc
     3781 tatgagcagc tcattggaaa actgatggat ggtgaaggag atccagattc gccgctctgg
     3841 aggcacccca tgctgtgcta cagcaaagac ggcctatacg cctccctcac caccctgccc
     3901 tctgaggcct tgcagacgga ggccctcaag ctcttcaagt cctgccagct cttcatcaac
     3961 gtgccggtgg aagctgcctc ggtggactac catgtgtccc tggcccagac cgcactgcag
     4021 gtctgcctgg ttcaccccga gctgcagagt gagatctact gccaactcat gaagcagacc
     4081 agctgccgcc cacctcagaa gtactccctc atgcagtgct ggcagctcct cgctctgtgt
     4141 gctccacttt tcctgcctca gcatcacttc ctctggtatg tcaagcagca gctccaacgc
     4201 catgcagatc ccagaagtga aactggccag tatgccacct actgccagcg ggcagtggag
     4261 cggaccctgc ggaccgggga gcgggaagcc aggccatcgc gcatggaagt ggtgtccatc
     4321 ctgctgcgta accccttcca ccactccttg cccttcagca tccccgtgca ctttaccaac
     4381 gggacttacc atgtggttgg ttttgacggc tcttccacgg ttgatgagtt cctccagcgg
     4441 ctgaaccagg agataggcat gagaaagcca tcccactctg gctttgccct cttcacggac
     4501 gatccctcgg gcagggacct ggagcactgc ctgcagggaa gtgtcaagat ctgtgatgcc
     4561 atctccaagt gggaacaagc catgaaggag ctgcaccccg gaaagtctga gggtgggaca
     4621 cgcgtcgtga agctgatgta caagaacagg ctgtactttc gcagtcaagt caaaggggag
     4681 acggaccgag aacggctgct ccttgcctct caaaccagta gagagatagt ggcagggagg
     4741 tttcctatca acaaggaatt ggctcttgag atggctgccc tgatggccca ggtagaatat
     4801 ggggacttgg agaagcctgc cctgccaggc cctggaggca catcccctgc caaggctcag
     4861 catcttctcc agcaggtcct agacaggttc caccccaggc gctatagaca tggggccccc
     4921 gctgaacagc tgaggcacct ggcagatatg ttgaccacaa aatgggcaac attgcaagga
     4981 tgctcccctc ctgagtgcat ccgcatctac ctgaccgtgg ccaggaaatg gcctttcttt
     5041 ggtgctaaac tttttgctgc tcagcctgcc cagctgtctt ccaaggagaa cgctctggtg
     5101 tggattgctg tgaatgagga tggcgtcagc atcctggacc acaacactat gcaagtgcac
     5161 atcacttacc cctactcttc agtgacaacg tttggtggct gcagggatga cttcatgctt
     5221 gtgattagat ctatcccaga caagagctct ggaaaaagcc acattgagaa gttgatcttc
     5281 cggatggctg ctcccaagat tgcagaagcc accttcatca tggccagcta tatgaaccat
     5341 tgcactacaa ctgtgaaccc ccccaccaac ccacccggag cctgccagct gtgggaactg
     5401 gatggacgac agttcttttc ttctgtttcc tgtgctacca aggggccaac gttgctgtga
     5461 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     5521 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     5581 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     5641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5701 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     5761 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5821 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5881 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5941 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6001 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6061 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6121 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6181 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6241 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6301 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     6361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6601 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6661 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     6721 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     6781 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     6841 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     6901 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     6961 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     7021 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     7081 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     7141 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     7201 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     7261 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     7321 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     7381 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     7441 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     7501 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     7561 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7621 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7681 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     7741 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     7801 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     7861 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     7921 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     7981 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8041 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8101 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8161 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8221 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     8281 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8341 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8401 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8461 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8521 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8581 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8641 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8701 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8761 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     8821 cat
//
