LOCUS       sequence-364710-        4828 bp    DNA     circular SYN 14-APR-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 Apr 14, 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_N1
FEATURES             Location/Qualifiers
     source          1..4828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      783..792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             789..1484
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(922..940)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1190..1209
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1609..1730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1646..1665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1700..1719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1737..2192)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1874..1895)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2086..2105
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2219..2323
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2325..2682
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2325..2516
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2348..2368)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2533..2668
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2717..3511
                     /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             2717..3511
                     /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(2771..2790)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3381..3400
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3699..3718)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3743..3790
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4119..4707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4608..4627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctggt accccttgca gcttccagtt gtcaggaaga gaaggctccc atggatcctg
      661 tggtggtgct ggggctctgt ctctcctgtt tgcttctcct ttcactctgg aaacagagct
      721 atgggggagg gaagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg
      781 tcgccaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
      841 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      901 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
      961 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg
     1021 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     1081 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     1141 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     1201 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     1261 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     1321 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     1381 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     1441 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagcg gccgcgactc
     1501 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1561 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1621 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1681 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     1741 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     1801 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     1861 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     1921 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     1981 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2041 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2101 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2161 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2221 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2281 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2341 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2401 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2461 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2521 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2581 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2641 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2701 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2761 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2821 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2881 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2941 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3001 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3061 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3121 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3181 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3241 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3301 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3361 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3421 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3481 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3541 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3601 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3661 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3721 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     3781 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     3841 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     3901 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     3961 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4021 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4081 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4141 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4201 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4261 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4321 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4381 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4441 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4501 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4561 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4621 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4681 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4741 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4801 aaccgtatta ccgccatgca ttagttat
//
