LOCUS       sequence-404744-        5580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses two repeats of PLEKHA5 aa 143-271 (K163A and R164A) fused 
            to mScarlet-iin mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  TITLE     A phosphorylation-controlled switch confers cell cycle-dependent 
            protein relocalization
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5580)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 2xMARS
FEATURES             Location/Qualifiers
     source          1..5580
                     /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      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1505..1514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1511..2206
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(1644..1662)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1912..1931
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2213..2239
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    2368..2489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2405..2424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2459..2478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2496..2951)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2633..2654)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2845..2864
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2978..3082
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3084..3441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3084..3275
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3107..3127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3292..3427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3476..4270
                     /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             3476..4270
                     /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(3530..3549)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4140..4159
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4458..4477)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4502..4549
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4878..5466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5367..5386
                     /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 ccggactcag atctcgagct caagcttcga attcgccacc atgggcagaa cttcacgagc
      661 ttcaaaaaaa gttcataatt ttggaaagag gtcaaattca attgcagcga atcctaatgc
      721 accggttgtc agacgaggtt ggctttataa acaggacagt actggcatga aattgtggaa
      781 gaaacgctgg tttgtgcttt ctgacctttg cctcttttat tatagagatg agaaagaaga
      841 gggtatcctg ggaagcatac tgttacctag ttttcagata gctttgctta cctctgaaga
      901 tcacattaat cgcaaatatg cttttaaggc agcccatcca aacatgcgga cctattattt
      961 ctgcactgat acaggaaagg aaatggagtt gtggatgaaa gccatgttag atgctgccct
     1021 agtacagaca ctgcagtcga cggtaccgag cggcggccgc ggatcaggaa gcggatcagg
     1081 aagcggaatg ggcagaactt cacgagcttc aaaaaaagtt cataattttg gaaagaggtc
     1141 aaattcaatt gcagcgaatc ctaatgcacc ggttgtcaga cgaggttggc tttataaaca
     1201 ggacagtact ggcatgaaat tgtggaagaa acgctggttt gtgctttctg acctttgcct
     1261 cttttattat agagatgaga aagaagaggg tatcctggga agcatactgt tacctagttt
     1321 tcagatagct ttgcttacct ctgaagatca cattaatcgc aaatatgctt ttaaggcagc
     1381 ccatccaaac atgcggacct attatttctg cactgataca ggaaaggaaa tggagttgtg
     1441 gatgaaagcc atgttagatg ctgccctagt acagacagta ccgcgggccc gggatccacc
     1501 ggtcgccacc atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt
     1561 gcacatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     1621 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     1681 ctcctgggac atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc
     1741 cgccgacatc cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt
     1801 gatgaacttc gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg
     1861 caccctgatc tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat
     1921 gcagaagaag acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt
     1981 gctgaagggc gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga
     2041 cttcaagacc acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga
     2101 ccgcaagttg gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg
     2161 ctccgagggc cgccactcca ccggcggcat ggacgagctg tacaagctgc agctgcctcc
     2221 cctggagcgc ctgaccctgg actaaagcgg ccgcgactct agatcataat cagccatacc
     2281 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2341 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2401 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2461 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2521 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2581 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2641 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2701 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2761 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2821 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2881 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2941 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3001 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3061 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3121 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3181 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3241 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3361 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3421 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3481 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3541 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3601 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3661 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3721 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3781 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3841 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3901 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3961 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4021 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4081 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4141 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4201 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4261 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4321 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4381 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4441 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4501 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4561 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4621 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4681 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4741 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4801 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4861 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4921 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4981 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5041 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5101 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5161 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5221 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5281 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5341 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5401 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5461 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5521 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
