LOCUS       sequence-502459-        5700 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  Mitochondria-tethered construct for the RAPID-Release approach.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5700)
  AUTHORS   Apta-Smith MJ, Hernandez-Fernaud JR, Bowman AJ
  TITLE     Evidence for the nuclear import of histones H3.1 and H4 as monomers.
  JOURNAL   EMBO J. 2018 Oct 1;37(19):e98714. doi: 10.15252/embj.201798714. Epub
            2018 Sep 3.
  PUBMED    30177573
REFERENCE   2  (bases 1 to 5700)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 10, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J."; 
            date: "2018-10-1"; volume: "37(19):e98714. doi"; pages: " 
            10.15252/embj.201798714. Epub 2018 Sep 3"
COMMENT     Sequence Label: pCL1046
FEATURES             Location/Qualifiers
     source          1..5700
                     /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      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1021..1030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1027..1734
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1172..1190)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1440..1459
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1774..1794
                     /codon_start=1
                     /product="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site
                     "
                     /label=TVMV site
                     /translation="ETVRFQS"
     CDS             1804..1824
                     /codon_start=1
                     /product="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site
                     "
                     /label=TVMV site
                     /translation="ETVRFQS"
     polyA_signal    2488..2609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2525..2544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2579..2598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2616..3071)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2753..2774)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2965..2984
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3098..3202
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3204..3561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3204..3395
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3227..3247)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3412..3547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3596..4390
                     /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             3596..4390
                     /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(3650..3669)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4260..4279
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4578..4597)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4622..4669
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4998..5586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5487..5506
                     /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 gctagccacc
      601 atggcccgta ccaagcagac cgcccgtaaa tccaccggag ggaaggctcc ccgcaagcag
      661 ctggccacca aggcagccag gaagtccgct cctgctaccg gcggagtcaa gaaacctcac
      721 cgttaccggc ccggcacagt cgctctccgc gagatccgcc gctaccagaa atccaccgag
      781 ctgctcatcc gcaaactgcc tttccagcgc ctggtccggg agatcgctca ggacttcaag
      841 accgacctgc gcttccagag ctcggccgtt atggctctgc aggaggccag cgaggcttat
      901 ctggtcgctc tctttgagga caccaacctg tgcgccatcc acgccaagag ggtcaccatc
      961 atgcccaagg acatccagct ggcccgcaga atccgaggcg agagggctgg cggaccggtc
     1021 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     1081 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1141 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1201 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1261 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1321 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1381 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1441 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1501 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1561 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1621 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1681 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtactca
     1741 gatctcgagg gaggttccgg cgggagcgga ggagaaactg taagattcca aagcggtgga
     1801 tccgaaaccg tgcgctttca gagcggtggc gggggaagcg gcggatcagg tggcgcggga
     1861 ggtggctctg gaggtagcgg cggcgtggcg attctgtggc atgaaatgtg gcatgaaggc
     1921 ctggaagaag cgagccgcct gtattttggc gaacgcaacg tgaaaggcat gtttgaagtg
     1981 ctggaaccgc tgcatgcgat gatggaacgc ggcccgcaga ccctgaaaga aaccagcttt
     2041 aaccaggcgt atggccgcga tctgatggaa gcgcaggaat ggtgccgcaa atatatgaaa
     2101 agcggcaacg tgaaagatct gacccaggcg tgggatctgt attatcatgt gtttcgccgc
     2161 attggcggca ccggcggcag cggaggtacc ggcggtagcg ggggatcagg tggcagcgga
     2221 gggaccggtg gaagcgagct caagcttcgc ggcgatggcg aaccgagcgg cgtgccggtg
     2281 gcggtggtgc tgctgccggt gtttgcgctg accctggtgg cggtgtgggc gtttgtgcgc
     2341 tatcgcaaac agctgtaggg atccaccgga tctagataac tgatcataat cagccatacc
     2401 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2461 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2521 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2581 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     2641 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2701 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2761 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2821 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2881 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2941 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3001 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3061 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3121 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3181 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3241 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3481 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3541 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3601 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3661 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3721 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3781 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3841 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3901 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3961 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4021 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4081 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4141 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4201 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4261 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4321 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4381 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4441 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4501 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4561 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4621 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4681 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4741 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4801 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4861 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4921 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4981 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5101 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5161 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5221 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5281 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5341 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5401 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5461 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5521 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5581 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5641 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
