LOCUS       sequence-358010-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the plasma membrane (can be used for 
            colocalization studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  AUTHORS   Chertkova AO, Mastop M, Postma M, van Bommel N, van der Niet S, 
            Batenburg KL, Joosen L, Gadella TWJ, Okada Y, Goedhart J
  TITLE     Robust and Bright Genetically Encoded Fluorescent Markers for 
            Highlighting Structures and Compartments in Mammalian Cells
  JOURNAL   bioRxiv (2020) 160374; doi: https://doi.org/10.1101/160374
REFERENCE   2  (bases 1 to 4743)
  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: "bioRxiv 
            (2020) 160374; doi: https://doi.org/10.1101/160374"
COMMENT     Sequence Label: Lck-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..4743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     284..304
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(329..433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      460..915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(547..566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     757..778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    922..1043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(933..952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     987..1006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    1167..1232
                     /label=MCS
                     /note="multiple cloning site"
     CDS             complement(1233..1928)
                     /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(1508..1527)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1777..1795
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1925..1934
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1970..1979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2006..2209)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2035..2055)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2210..2513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2688..3276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2768..2787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3605..3652
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3677..3696
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3884..4678)
                     /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             complement(3884..4678)
                     /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(3995..4014)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4605..4624
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4713..4743,1..327))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4727..4743,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
       61 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
      121 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
      181 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      241 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      301 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
      361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
      481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      901 taacgcttac aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa
      961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1021 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1081 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     1141 attatgatca gttatctaga tccggtggat cccgggcccg cggtaccgtc gactgcagaa
     1201 ttcgaagctt gagctcgaga tctgagtccg gacttgtaca gctcgtccat gccgccggtg
     1261 gagtggcggc cctcggagcg ttcgtactgt tccaccacgg tgtagtcctc gttgtgggag
     1321 gtgatgtcca acttgcggtc gacgttgtag gcgccgggca tctgcacggg cttcttggcc
     1381 ttgtaggtgg tcttgaagtc cgccaggtag cggccgccgt ccttcaggcg cagggccatc
     1441 ttaatgtcgc ccttcagcac gccgtcctcg gggtacaacc gctcggtgga cgcttcccag
     1501 cccattgtct tcttctgcat tacggggccg tcaggaggga agttggtgcc gcggagcttc
     1561 accttgtaga tcagggtgcc gtcctccagg gaggtgtcct gggtcacggt cacggcgccg
     1621 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga ctgcttatag
     1681 tagtcgggga tgtcggcggg gtgcttgatg aaggccctgg agccgtacat gaactgaggg
     1741 gacaggatgt cccaggagaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc
     1801 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     1861 atggagccct ccatgtgcac cttgaaccgc atgaactcct tgatcactgc ctcgcccttg
     1921 ctcaccatgg tggcgaccgg tagctcgggg ttgctgctgc acacgcagcc catggtggct
     1981 agcggatctg acggttcact aaaccagctc tgcttatata gacctcccac cgtacacgcc
     2041 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     2101 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     2161 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     2221 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     2281 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     2341 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     2401 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     2461 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     2521 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctaatgcatg
     2581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3121 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3421 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     3481 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     3541 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     3601 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     3661 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     3721 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     3781 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     3841 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     3901 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     3961 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4021 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4081 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4141 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4201 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     4261 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     4321 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     4381 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     4441 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     4501 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     4561 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     4621 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     4681 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     4741 cct
//
