LOCUS       sequence-368865-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Plekhh1-NTer labelled with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  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 6348)
  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-NTer
FEATURES             Location/Qualifiers
     source          1..6348
                     /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    3136..3257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3173..3192)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3227..3246
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3264..3719)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3401..3422)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3613..3632
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3746..3850
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3852..4209
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3852..4043
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3875..3895)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4060..4195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4244..5038
                     /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             4244..5038
                     /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(4298..4317)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4908..4927
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5226..5245)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5270..5317
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5646..6234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6135..6154
                     /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 gttgagtcga cggtaccgcg
     3001 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     3061 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3121 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3181 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3241 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3301 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3361 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3421 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3481 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3541 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3601 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3661 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3721 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3781 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3841 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3961 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4081 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4141 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4201 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4261 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4321 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4381 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4441 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4501 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4561 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4621 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4681 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4741 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4801 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4861 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4921 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4981 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5041 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5101 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5161 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     5221 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5281 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5341 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5401 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5461 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5521 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5581 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5641 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5701 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5761 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5821 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5881 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5941 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6001 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6061 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6121 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6181 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6241 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6301 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
