LOCUS       sequence-368868-        6450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Plekhh1-MyTH-FERM labelled with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  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 6450)
  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-MyTH-FERM
FEATURES             Location/Qualifiers
     source          1..6450
                     /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    3238..3359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3275..3294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3329..3348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3366..3821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3503..3524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3715..3734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3848..3952
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3954..4311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3954..4145
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3977..3997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4162..4297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4346..5140
                     /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             4346..5140
                     /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(4400..4419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5010..5029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5328..5347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5372..5419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5748..6336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6237..6256
                     /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 attctacagt ggctgcaggt
     1381 ggcagcagtg ccaaggtggg cactgcctat gagcagctca ttggaaaact gatggatggt
     1441 gaaggagatc cagattcgcc gctctggagg caccccatgc tgtgctacag caaagacggc
     1501 ctatacgcct ccctcaccac cctgccctct gaggccttgc agacggaggc cctcaagctc
     1561 ttcaagtcct gccagctctt catcaacgtg ccggtggaag ctgcctcggt ggactaccat
     1621 gtgtccctgg cccagaccgc actgcaggtc tgcctggttc accccgagct gcagagtgag
     1681 atctactgcc aactcatgaa gcagaccagc tgccgcccac ctcagaagta ctccctcatg
     1741 cagtgctggc agctcctcgc tctgtgtgct ccacttttcc tgcctcagca tcacttcctc
     1801 tggtatgtca agcagcagct ccaacgccat gcagatccca gaagtgaaac tggccagtat
     1861 gccacctact gccagcgggc agtggagcgg accctgcgga ccggggagcg ggaagccagg
     1921 ccatcgcgca tggaagtggt gtccatcctg ctgcgtaacc ccttccacca ctccttgccc
     1981 ttcagcatcc ccgtgcactt taccaacggg acttaccatg tggttggttt tgacggctct
     2041 tccacggttg atgagttcct ccagcggctg aaccaggaga taggcatgag aaagccatcc
     2101 cactctggct ttgccctctt cacggacgat ccctcgggca gggacctgga gcactgcctg
     2161 cagggaagtg tcaagatctg tgatgccatc tccaagtggg aacaagccat gaaggagctg
     2221 caccccggaa agtctgaggg tgggacacgc gtcgtgaagc tgatgtacaa gaacaggctg
     2281 tactttcgca gtcaagtcaa aggggagacg gaccgagaac ggctgctcct tgcctctcaa
     2341 accagtagag agatagtggc agggaggttt cctatcaaca aggaattggc tcttgagatg
     2401 gctgccctga tggcccaggt agaatatggg gacttggaga agcctgccct gccaggccct
     2461 ggaggcacat cccctgccaa ggctcagcat cttctccagc aggtcctaga caggttccac
     2521 cccaggcgct atagacatgg ggcccccgct gaacagctga ggcacctggc agatatgttg
     2581 accacaaaat gggcaacatt gcaaggatgc tcccctcctg agtgcatccg catctacctg
     2641 accgtggcca ggaaatggcc tttctttggt gctaaacttt ttgctgctca gcctgcccag
     2701 ctgtcttcca aggagaacgc tctggtgtgg attgctgtga atgaggatgg cgtcagcatc
     2761 ctggaccaca acactatgca agtgcacatc acttacccct actcttcagt gacaacgttt
     2821 ggtggctgca gggatgactt catgcttgtg attagatcta tcccagacaa gagctctgga
     2881 aaaagccaca ttgagaagtt gatcttccgg atggctgctc ccaagattgc agaagccacc
     2941 ttcatcatgg ccagctatat gaaccattgc actacaactg tgaacccccc caccaaccca
     3001 cccggagcct gccagctgtg ggaactggat ggacgacagt tcttttcttc tgtttcctgt
     3061 gctaccaagg ggccaacgtt gctgtgagtc gacggtaccg cgggcccggg atccaccgga
     3121 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3181 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3241 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3301 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3361 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3421 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3481 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3541 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3601 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3661 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3721 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3781 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3841 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3901 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3961 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4141 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4201 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4261 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4321 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4381 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4441 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4501 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4561 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4621 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4681 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4741 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4801 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4861 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4921 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4981 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5041 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5101 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5161 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5221 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5281 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5341 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5401 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5461 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5521 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5581 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5641 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5701 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5761 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5821 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5881 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5941 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6001 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6061 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6121 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6181 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6241 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6301 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6361 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6421 tctgtggata accgtattac cgccatgcat
//
