LOCUS       sequence-163908-        6484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Merlin with an alanine mutation at the Pak 
            phosphorylation site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6484)
  AUTHORS   Xiao GH, Beeser A, Chernoff J, Testa JR
  TITLE     p21-activated kinase links Rac/Cdc42 signaling to merlin.
  JOURNAL   J Biol Chem. 2002 Jan 11. 277(2):883-6.
  PUBMED    11719502
REFERENCE   2  (bases 1 to 6484)
  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 Biol 
            Chem. 2002 Jan 11. 277(2):883-6."
COMMENT     Sequence Label: pEGFP-Merlin S518A
FEATURES             Location/Qualifiers
     source          1..6484
                     /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    3272..3393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3309..3328)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3363..3382
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3400..3855)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3537..3558)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3749..3768
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3882..3986
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3988..4345
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3988..4179
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4011..4031)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4196..4331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4380..5174
                     /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             4380..5174
                     /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(4434..4453)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5044..5063
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5362..5381)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5406..5453
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5782..6370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6271..6290
                     /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 atccgccggg gccatcgctt cccgcatgag cttcagctct
     1381 ctcaagagga agcaacccaa gacgttcacc gtgaggatcg tcaccatgga cgccgagatg
     1441 gagttcaatt gcgagatgaa gtggaaaggg aaggacctct ttgatttggt gtgccggact
     1501 ctggggctcc gagaaacctg gttctttgga ctgcagtaca caatcaagga cacagtggcc
     1561 tggctcaaaa tggacaagaa ggtactggat catgatgttt caaaggaaga accagtcacc
     1621 tttcacttct tggccaaatt ttatcctgag aatgctgaag aggagctggt tcaggagatc
     1681 acacaacatt tattcttctt acaggtaaag aagcagattt tagatgaaaa gatctactgc
     1741 cctcctgagg cttctgtgct cctggcttct tacgccgtcc aggccaagta tggtgactac
     1801 gaccccagtg ttcacaagcg gggatttttg gcccaagagg aattgcttcc aaaaagggta
     1861 ataaatctgt atcagatgac tccggaaatg tgggaggaga gaattactgc ttggtacgca
     1921 gagcaccgag gccgagccag ggatgaagct gaaatggaat atctgaagat agctcaggac
     1981 ctggagatgt acggtgtgaa ctactttgca atccggaata aaaagggcac agagctgctg
     2041 cttggagtgg atgccctggg gcttcacatt tatgaccctg agaacagact gacccccaag
     2101 atctccttcc cgtggaatga aatccgaaac atctcgtaca gtgacaagga gtttactatt
     2161 aaaccactgg ataagaaaat tgatgtcttc aagtttaact cctcaaagct tcgtgttaat
     2221 aagctgattc tccagctatg tatcgggaac catgatctat ttatgaggag aaggaaagcc
     2281 gattctttgg aagttcagca gatgaaagcc caggccaggg aggagaaggc tagaaagcag
     2341 atggagcggc agcgcctcgc tcgagagaag cagatgaggg aggaggctga acgcacgagg
     2401 gatgagttgg agaggaggct gctgcagatg aaagaagaag caacaatggc caacgaagca
     2461 ctgatgcggt ctgaggagac agctgacctg ttggctgaaa aggcccagat caccgaggag
     2521 gaggcaaaac ttctggccca gaaggccgca gaggctgagc aggaaatgca gcgcatcaag
     2581 gccacagcga ttcgcacgga ggaggagaag cgcctgatgg agcagaaggt gctggaagcc
     2641 gaggtgctgg cactgaagat ggctgaggag tcagagagga gggccaaaga ggcagatcag
     2701 ctgaagcagg acctgcagga agcacgcgag gcggagcgaa gagccaagca gaagctcctg
     2761 gagattgcca ccaagcccac gtacccgccc atgaacccaa ttccagcacc gttgcctcct
     2821 gacataccaa gcttcaacct cattggtgac agcctgtctt tcgacttcaa agatactgac
     2881 atgaagcggc ttgccatgga gatagagaaa gaaaaagtgg aatacatgga aaagagcaag
     2941 catctgcagg agcagctcaa tgaactcaag acagaaatcg aggccttgaa actgaaagag
     3001 agggagacag ctctggatat tctgcacaat gagaactccg acaggggtgg cagcagcaag
     3061 cacaatacca ttaaaaagct caccttgcag agcgccaagt cccgagtggc cttctttgaa
     3121 gagctctagt ctagagggcc cgggatccac cggatctaga taactgatca taatcagcca
     3181 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3241 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3361 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     3421 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3481 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3541 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3601 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3661 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3721 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3781 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3841 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3901 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3961 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4021 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4081 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4141 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4201 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4261 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4321 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4381 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4441 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4501 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4561 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4621 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4681 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4741 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4801 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4861 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4921 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4981 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5041 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5101 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5161 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5221 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5281 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5341 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5401 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5461 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5521 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5581 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6481 gcat
//
