LOCUS       sequence-368874-        5913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Plekhh1-FERM labelled with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5913)
  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 5913)
  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-FERM
FEATURES             Location/Qualifiers
     source          1..5913
                     /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    2701..2822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2738..2757)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2792..2811
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2829..3284)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2966..2987)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3178..3197
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3311..3415
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3417..3774
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3417..3608
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3440..3460)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3625..3760
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3809..4603
                     /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             3809..4603
                     /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(3863..3882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4473..4492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4791..4810)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4835..4882
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5211..5799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5700..5719
                     /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 attctgggga gcgggaagcc
     1381 aggccatcgc gcatggaagt ggtgtccatc ctgctgcgta accccttcca ccactccttg
     1441 cccttcagca tccccgtgca ctttaccaac gggacttacc atgtggttgg ttttgacggc
     1501 tcttccacgg ttgatgagtt cctccagcgg ctgaaccagg agataggcat gagaaagcca
     1561 tcccactctg gctttgccct cttcacggac gatccctcgg gcagggacct ggagcactgc
     1621 ctgcagggaa gtgtcaagat ctgtgatgcc atctccaagt gggaacaagc catgaaggag
     1681 ctgcaccccg gaaagtctga gggtgggaca cgcgtcgtga agctgatgta caagaacagg
     1741 ctgtactttc gcagtcaagt caaaggggag acggaccgag aacggctgct ccttgcctct
     1801 caaaccagta gagagatagt ggcagggagg tttcctatca acaaggaatt ggctcttgag
     1861 atggctgccc tgatggccca ggtagaatat ggggacttgg agaagcctgc cctgccaggc
     1921 cctggaggca catcccctgc caaggctcag catcttctcc agcaggtcct agacaggttc
     1981 caccccaggc gctatagaca tggggccccc gctgaacagc tgaggcacct ggcagatatg
     2041 ttgaccacaa aatgggcaac attgcaagga tgctcccctc ctgagtgcat ccgcatctac
     2101 ctgaccgtgg ccaggaaatg gcctttcttt ggtgctaaac tttttgctgc tcagcctgcc
     2161 cagctgtctt ccaaggagaa cgctctggtg tggattgctg tgaatgagga tggcgtcagc
     2221 atcctggacc acaacactat gcaagtgcac atcacttacc cctactcttc agtgacaacg
     2281 tttggtggct gcagggatga cttcatgctt gtgattagat ctatcccaga caagagctct
     2341 ggaaaaagcc acattgagaa gttgatcttc cggatggctg ctcccaagat tgcagaagcc
     2401 accttcatca tggccagcta tatgaaccat tgcactacaa ctgtgaaccc ccccaccaac
     2461 ccacccggag cctgccagct gtgggaactg gatggacgac agttcttttc ttctgtttcc
     2521 tgtgctacca aggggccaac gttgctgtga gtcgacggta ccgcgggccc gggatccacc
     2581 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2641 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2701 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2761 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2821 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2881 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2941 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3001 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3061 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3121 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3181 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3241 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3301 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3361 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3421 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3481 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3541 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3601 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3661 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3721 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3781 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3841 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3901 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3961 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4021 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4081 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4141 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4201 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4261 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4321 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4381 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4441 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4501 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4561 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4621 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4681 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4741 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4801 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4861 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4921 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4981 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5041 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5101 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5161 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5221 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5281 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5341 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5401 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5461 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5521 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5581 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5641 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5701 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5761 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5821 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5881 gattctgtgg ataaccgtat taccgccatg cat
//
