LOCUS       sequence-359747-        5086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization of mScarlet-I fluorescent protein to histone 2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5086)
  AUTHORS   Degrelle SA, Ferecatu I, Fournier T
  TITLE     Novel fluorescent and secreted transcriptional reporters for 
            quantifying activity of the xenobiotic sensor aryl hydrocarbon 
            receptor (AHR).
  JOURNAL   Environ Int. 2022 Sep 24;169:107545. doi: 
            10.1016/j.envint.2022.107545.
  PUBMED    36179647
REFERENCE   2  (bases 1 to 5086)
  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: "Environ 
            Int."; date: "2022-09-24"; volume: "169:107545. doi"; pages: " 
            10.1016/j.envint.2022.107545"
COMMENT     Sequence Label: CMV-H2B-mScarlet-I
FEATURES             Location/Qualifiers
     source          1..5086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        94..285
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     242..262
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(287..391)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      418..873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(505..524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     715..736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    880..1001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(891..910)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     945..964
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1127..1822)
                     /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(1402..1421)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1671..1689
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1819..1828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1841..2218)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     promoter        complement(2307..2510)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2336..2356)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2511..2814
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2989..3577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3069..3088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3906..3953
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3978..3997
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4185..4979)
                     /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(4185..4979)
                     /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(4296..4315)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4906..4925
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5014..5086,1..285))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5028..5086,1..77)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
       61 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
      121 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
      181 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      241 caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc
      301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
      361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      421 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      481 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      541 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      601 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      661 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      721 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      781 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      841 cgaattttaa caaaatatta acgcttacaa tttacgcctt aagatacatt gatgagtttg
      901 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      961 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1021 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     1081 acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg tacagctcgt
     1141 ccatgccgcc ggtggagtgg cggccctcgg agcgttcgta ctgttccacc acggtgtagt
     1201 cctcgttgtg ggaggtgatg tccaacttgc ggtcgacgtt gtaggcgccg ggcatctgca
     1261 cgggcttctt ggccttgtag gtggtcttga agtccgccag gtagcggccg ccgtccttca
     1321 ggcgcagggc catcttaatg tcgcccttca gcacgccgtc ctcggggtac aaccgctcgg
     1381 tggacgcttc ccagcccatt gtcttcttct gcattacggg gccgtcagga gggaagttgg
     1441 tgccgcggag cttcaccttg tagatcaggg tgccgtcctc cagggaggtg tcctgggtca
     1501 cggtcacggc gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     1561 aggactgctt atagtagtcg gggatgtcgg cggggtgctt gatgaaggcc ctggagccgt
     1621 acatgaactg aggggacagg atgtcccagg agaagggcag ggggccaccc ttggtcacct
     1681 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     1741 actcgtggcc gttcatggag ccctccatgt gcaccttgaa ccgcatgaac tccttgatca
     1801 ctgcctcgcc cttgctcacc atggtggcga ccggtggatc cttagcgctg gtgtacttgg
     1861 tgatggcctt agtaccctcg gacacggcgt gcttggccaa ctccccaggc agcagcaggc
     1921 gcacggccgt ctggatctcc ctggaggtga tggtcgagcg cttgttgtaa tgcgccaggc
     1981 gggaagcctc acctgcgatg cgctcgaaaa tgtcgttcac aaacgaattc atgatgccca
     2041 tggccttgga cgaaatgccg gtgtcagggt ggacctgctt cagaaccttg tacacataga
     2101 tggaatagct ctccttgcgg ctgcgcttgc gcttcttgcc gcctttcttc tgcgccttag
     2161 tcaccgcctt cttggagccc tttttcgggg cgggagcaga cttcgctggc tctggcatgg
     2221 tggcggtacc gtcgactgca gaattcgaag cttgagctcg agatctgagt ccggtagcgc
     2281 tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca ccgtacacgc
     2341 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     2401 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     2461 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     2521 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     2581 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     2641 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     2701 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     2761 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     2821 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actaatgcat
     2881 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2941 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3001 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3061 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3121 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3181 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3241 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3301 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3361 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3421 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3481 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3541 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3601 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3661 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3721 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
     3781 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
     3841 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
     3901 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
     3961 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
     4021 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     4081 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     4141 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     4201 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     4261 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     4321 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     4381 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     4441 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     4501 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     4561 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     4621 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     4681 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     4741 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcttgc
     4801 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     4861 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     4921 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     4981 cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa
     5041 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcg
//
