LOCUS       sequence-163859-        5075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of the nucleus (can be used for colocalization
            studies).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5075)
  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 5075)
  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: pmScarlet-i_H2A_C1
FEATURES             Location/Qualifiers
     source          1..5075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     158..177
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        291..395
                     /gene="bla"
                     /label=AmpR promoter
     promoter        397..754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        397..588
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(420..440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      605..740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             789..1583
                     /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             789..1583
                     /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(843..862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1453..1472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1771..1790)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1815..1862
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2191..2779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2680..2699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2954..3257
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3258..3461
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3412..3432
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3500..3509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3506..4201
                     /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(3639..3657)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3907..3926
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4756..4877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4793..4812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4847..4866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4884..5075,1..264))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5021..5042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
       61 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
      121 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
      181 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
      241 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      361 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
      421 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      481 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      601 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      661 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      721 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
      781 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
      841 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
      901 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
      961 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     1021 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     1081 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     1141 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     1201 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     1261 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     1321 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     1381 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     1441 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     1501 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     1561 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     1621 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     1681 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1741 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     1801 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     1861 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     1921 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     1981 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     2041 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     2101 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2161 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2221 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2281 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2341 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2401 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2461 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2521 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2581 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2641 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2701 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2761 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2821 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2881 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     2941 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3001 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3061 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3121 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3181 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3241 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     3301 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3361 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3421 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     3481 tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggcagtga tcaaggagtt
     3541 catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg agatcgaggg
     3601 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     3661 tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg gctccagggc
     3721 cttcatcaag caccccgccg acatccccga ctactataag cagtccttcc ccgagggctt
     3781 caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga cccaggacac
     3841 ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca acttccctcc
     3901 tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcatccaccg agcggttgta
     3961 ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga aggacggcgg
     4021 ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc agatgcccgg
     4081 cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact acaccgtggt
     4141 ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     4201 gtactcagat ctcgagctca agcttcgaat tctgatgtcg ggacgcggca agcagggagg
     4261 caaagctcgc gccaaagcca agacccgctc ttctcgtgcc ggtctccagt tccccgtggg
     4321 ccgagtgcac cgactgctcc gcaagggcaa ctatgctgag cgggtcgggg ccggcgcgcc
     4381 ggtgtacctg gcggcggtgc tggagtacct gactgccgag atcctggagc tggcgggcaa
     4441 cgccgcccgc gacaacaaga agacccgcat tatcccgcgc cacttgcagc tggccatccg
     4501 caacgacgag gagctcaaca agctgctggg caaagtaacc atcgctcagg gtggtgtcct
     4561 gcccaacatc caggctgtgc tactgcccaa gaagaccgag agtcaccaca aggccaaagg
     4621 caaacgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     4681 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4741 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4801 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4861 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     4921 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4981 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5041 actattaaag aacgtggact ccaacgtcaa agggc
//
