LOCUS       sequence-163874-        4828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the ER (can be used for colocalization 
            studies and the OSER assay).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4828)
  AUTHORS   Bindels DS, Haarbosch L, van Weeren L, Postma M, Wiese KE, Mastop M,
            Aumonier S, Gotthard G, Royant A, Hink MA, Gadella TW Jr
  TITLE     mScarlet: a bright monomeric red fluorescent protein for cellular 
            imaging.
  JOURNAL   Nat Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074.
  PUBMED    27869816
REFERENCE   2  (bases 1 to 4828)
  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: "Nat 
            Methods. 2016 Nov 21. doi: 10.1038/nmeth.4074."
COMMENT     Sequence Label: pCytERM_mScarlet-i_N1
FEATURES             Location/Qualifiers
     source          1..4828
                     /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      790..799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             796..1491
                     /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(929..947)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1197..1216
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1616..1737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1653..1672)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1707..1726
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1744..2199)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1881..1902)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2093..2112
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2226..2330
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2332..2689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2332..2523
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2355..2375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2540..2675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2724..3518
                     /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             2724..3518
                     /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(2778..2797)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3388..3407
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3706..3725)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3750..3797
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4126..4714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4615..4634
                     /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 atctggtacc ccttgcagct tccagttgtc aggaagagaa ggctcccatg
      661 gatcctgtgg tggtgctggg gctctgtctc tcctgtttgc ttctcctttc actctggaaa
      721 cagagctatg ggggagggaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
      781 ccaccggtcg ccaccatggt gagcaagggc gaggcagtga tcaaggagtt catgcggttc
      841 aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
      901 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
      961 cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc cttcatcaag
     1021 caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt caagtgggag
     1081 cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac ctccctggag
     1141 gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc tgacggcccc
     1201 gtaatgcaga agaagacaat gggctgggaa gcatccaccg agcggttgta ccccgaggac
     1261 ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg ccgctacctg
     1321 gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg cgcctacaac
     1381 gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt ggaacagtac
     1441 gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagcggcc
     1501 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     1561 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     1621 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1681 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     1741 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1801 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1861 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1921 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1981 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     2041 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     2101 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     2161 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     2221 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2281 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     2341 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2401 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2461 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2521 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2581 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2641 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     2701 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     2761 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2821 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2881 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     2941 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3001 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3061 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3121 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3181 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3241 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3301 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3361 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3421 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3481 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3541 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     3601 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3661 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     3721 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     3781 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     3841 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     3901 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3961 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4021 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4081 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4201 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4261 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4321 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4381 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4441 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4501 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4561 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4621 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4681 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4741 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4801 tgtggataac cgtattaccg ccatgcat
//
