LOCUS       sequence-388852-        8199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express eGFP-RAPH1 lacking the TBS domain (RAPH1 aa 2-92 deleted) in
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8199)
  AUTHORS   Popovic A, Miihkinen M, Ghimire S, Saup R, Gronloh MLB, Ball NJ, 
            Goult BT, Ivaska J, Jacquemet G
  TITLE     Myosin-X recruits lamellipodin to filopodia tips.
  JOURNAL   J Cell Sci. 2023 Mar 1;136(5):jcs260574. doi: 10.1242/jcs.260574. 
            Epub 2023 Mar 2.
  PUBMED    36861887
REFERENCE   2  (bases 1 to 8199)
  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: "2023-03-1"; volume: "136(5):jcs260574. doi"; pages: " 
            10.1242/jcs.260574. Epub 2023 Mar 2"
COMMENT     Sequence Label: GFP-RAPH1-Delta-TBS
FEATURES             Location/Qualifiers
     source          1..8199
                     /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"
     regulatory      4660..4669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4987..5108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5024..5043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5078..5097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5115..5570)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5252..5273)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5464..5483
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5597..5701
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5703..6060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5703..5894
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5726..5746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5911..6046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6095..6889
                     /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             6095..6889
                     /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(6149..6168)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6759..6778
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7077..7096)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7121..7168
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7497..8085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7986..8005
                     /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 atctcgagcc gcgatgtgct ctatagagca ggagctcagc
     1381 agcattggtt caggaaacag taagcgtcaa atcacagaaa cgaaagctac tcagaaattg
     1441 cctgttagcc gacatacatt gaaacatggc accttgaaag gattatcttc ttcatctaat
     1501 aggatagcta aaccttccca tgccagctac tccttggacg acgtcactgc acagttagaa
     1561 caggcctctt tgagtatgga tgaggctgct cagcaatctg tactagaaga tactaaaccc
     1621 ttagtaacta atcagcacag aagaaccgcg tcagcaggca cagtgagtga tgctgaagta
     1681 cactctatta gtaattcctc ccattccagc atcacttccg cagcctccag catggactct
     1741 ttggatattg ataaagtaac acgccctcaa gagctggatt tgacacatca agggcagcca
     1801 attactgagg aagaacaggc agcaaaattg aaagctgaga agatcagagt tgccctagag
     1861 aaaattaaag aggcacaagt gaaaaagctg gtgatcagag tccacatgtc tgatgacagt
     1921 tctaaaacaa tgatggtgga tgagaggcag acagtaagac aagtactgga taacctgatg
     1981 gacaaatccc actgcggtta tagtttagac tggtcactgg tagaaaccgt ttctgaatta
     2041 caaatggaga gaatctttga agaccatgaa aacttggttg aaaatcttct taattggaca
     2101 agagatagcc aaaacaagct tatatttatg gagcgtatag aaaaatatgc acttttcaaa
     2161 aacccacaga attatctttt ggggaaaaag gaaacagctg agatggcaga tagaaacaaa
     2221 gaagtcctct tggaggaatg tttttgtgga agttctgtaa ctgtaccaga aattgaagga
     2281 gtcctttggt tgaaggatga tggcaagaag tcctggaaaa agcgttattt tctcttgcga
     2341 gcatctggta tctactatgt tcccaaagga aaagcaaagg tctctcggga tctggtgtgc
     2401 tttctccagc tggatcatgt caacgtttat tatggccagg actatcggaa caaatacaaa
     2461 gcacctacag actattgtct ggtgctgaag catccacaaa tccagaagaa atctcaatat
     2521 atcaaatacc tttgttgtga tgatgtgagg acactgcatc agtgggtcaa tgggatccgc
     2581 attgcaaagt atgggaagca gctctatatg aactaccaag aagccttgaa gaggacagag
     2641 tcagcctatg attggacttc cttatccagc tccagcatta aatcgggatc cagttcttcc
     2701 agcatcccag agtctcagtc aaaccactcc aatcagtctg atagcggagt ttctgacacc
     2761 cagccagcag gacacgtccg ttcccagagc attgtgagct ccgtattctc tgaagcctgg
     2821 aaacgaggca ctcagttgga agagtccagc aaggccagaa tggagtctat gaatcggccc
     2881 tacacttcac ttgtgccccc tttatccccg caacctaaga tagtcacccc ctacactgct
     2941 tcacagcctt caccacctct acctcctccg ccacccccac ctcctcctcc accaccccct
     3001 ccaccacccc ctcctccccc actccccagc cagtctgcac cttctgcagg ctcagcagcc
     3061 ccaatgttcg tcaagtacag cacaataaca cggctacaga atgcgtctca gcattcaggg
     3121 gccctgttta agccgccaac acccccagtg atgcagtcac agtcagtgaa gcctcagatc
     3181 ctggtacccc ccaatggagt tgttccacca ccccctcccc ctcctccacc cccaacccca
     3241 ggctctgcca tggcccagct aaagcctgca ccgtgtgccc catcccttcc acagttcagt
     3301 gccccgcctc ctccactgaa gatccatcaa gttcagcata ttactcaggt ggctccccca
     3361 acaccccccc cacctcctcc tatccctgca cccctccctc cccaagctcc cccaaaaccc
     3421 cttgtgacca tccccgcacc aaccagcacc aagactgtgg cacctgttgt gactcaagct
     3481 gcaccaccca cacctactcc tccagtgccc ccagcaaaaa agcagccagc tttccctgct
     3541 tcttacattc caccctctcc ccctacccct cctgttccag tacccccgcc aacattaccc
     3601 aagcaacaga gcttctgtgc aaaaccccct ccctctccac tgtcaccggt gccctcggtc
     3661 gtgaagcaga tagccagcca gtttccaccc cctccaactc cccctgccat ggaatctcag
     3721 cccttaaagc ctgtcccagc aaatgtagct ccacagtccc ctcctgcagt aaaagcaaag
     3781 cccaagtggc agcccagctc catcccagtc ccttctccgg acttccctcc tccccctcct
     3841 gaaagcagcc tggtgtttcc tcctccaccc ccatcacctg tcccagcccc accaccgcca
     3901 cctccaccca cagcttctcc tacccctgac aaaagtggat ctccaggcaa aaagaccagt
     3961 aagacgtcca gccctggggg aaagaaacca cccccaaccc cacagcgcaa ctccagcatt
     4021 aaatccagca gtggtgcaga gcaccccgag cccaagagac cctcggtgga cagtctagtc
     4081 agcaagttta caccgccagc agaatcaggg tctcccagca aggagaccct accacctcct
     4141 gcagcacccc ccaagcctgg aaaactcaat ctttctggag tcaaccttcc tggagttctc
     4201 caacaagggt gtgtgtcagc aaaagcccct gttctgagtg ggcgtggaaa ggactccgtg
     4261 gtggaatttc cttctcctcc atccgattct gattttccac cccctccacc tgaaacagac
     4321 cttcctctgc cccccattga gattccagca gttttctcgg gaaacacctc tccaaaagtg
     4381 gcagtcgtta atcctcaacc acaacaatgg tctaaaatgt cagtgaagaa ggcccctcca
     4441 cccacacgac ccaaacggaa tgatagcacc cgcctcactc aagctgagat ttctgagcag
     4501 ccaacaatgg ccacagttgt gccacaagtg cccacctctc ccaaatccag ccttagtgtc
     4561 cagcctggat tcctggctga cctcaacagg acactgcaac gaaagtccat cactcggcac
     4621 ggctcactct cctcccgcat gtccagagca gaaccaacag ccaccatgga tgatatggca
     4681 ttgcctccac caccccctga actgctgtct gatcaacaga aggctggtta cggaggcagt
     4741 catatatcag gctatgcaac gttgcggaga ggaccccctc ctgctccccc caaaagagac
     4801 cagaacacca agctctccag agactggtag ccaccatagg actttaccgc gggcccggga
     4861 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     4921 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4981 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5041 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5101 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     5161 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     5221 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     5281 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     5341 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     5401 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     5461 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5521 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     5581 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5641 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5701 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5761 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5821 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5881 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5941 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     6001 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     6061 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     6121 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     6181 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     6241 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     6301 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     6361 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     6421 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     6481 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6541 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     6601 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6661 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6721 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6781 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6841 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6901 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6961 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     7021 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     7081 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     7141 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     7201 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     7261 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     7321 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     7381 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7621 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8161 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
