LOCUS       sequence-373204-        7169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses HP1 with C-terminal tags ECFP and SNAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7169)
  AUTHORS   Vester K, Preussner M, Holton N, Feng S, Schultz C, Heyd F, Wahl MC
  TITLE     Recruitment of a splicing factor to the nuclear lamina for its 
            inactivation.
  JOURNAL   Commun Biol. 2022 Jul 22;5(1):736. doi: 10.1038/s42003-022-03689-y.
  PUBMED    35869234
REFERENCE   2  (bases 1 to 7169)
  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: "Commun 
            Biol."; date: "2022-07-22"; volume: "5(1):736. doi"; pages: " 
            10.1038/s42003-022-03689-y"
COMMENT     Sequence Label: HP1-ECFP-SNAP
FEATURES             Location/Qualifiers
     source          1..7169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1509..1518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1515..1517,1518..1520,1521..2231)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1560..1581)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1821..1840)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2168..2189
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2289..2834
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     intron          2945..3174
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3157..3176
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(3247..3821,3822..3824,3825..3833)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3414..3431)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3641..3660
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3856..4659
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3919..3938)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4529..4548
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4789..4910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4826..4845)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5012..5028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5012..5028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5025..5047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5036..5052
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5060..5066,5067..5084,5085..5090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5105..5126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5243..5260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5414..6002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5494..5513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6173..6964,6965..7033))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6796..6815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7034..7138)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgtcgctagc accatgggaa agaaaaccaa gcggacagct gacagttctt
      961 cttcagagga tgaggaggag tatgtcgtgg agaaggtgct agacaggcgc gtggttaagg
     1021 gacaagtgga atatctactg aagtggaaag gcttttctga ggagcacaat acttgggaac
     1081 ctgagaaaaa cttggattgc cctgagctaa tttctgaatt tatgaaaaag tataagaaga
     1141 tgaaggaggg tgaaaataat aaacccaggg agaagtcaga aagtaacaag aggaaatcca
     1201 atttctcaaa cagtgccgat gacatcaaat ctaaaaaaaa gagagagcag agcaatgata
     1261 tcgctcgggg ctttgagaga ggactggaac cagaaaagat cattggggca acagattcct
     1321 gtggtgattt aatgttccta atgaaatgga aagacacaga tgaagctgac ctggttcttg
     1381 caaaagaagc taatgtgaaa tgtccacaaa ttgtgatagc attttatgaa gagagactga
     1441 catggcatgc atatcctgag gatgcggaaa acaaagagaa agaaacagca aagagcggta
     1501 ccgagctcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1561 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1621 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1681 tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc
     1741 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1801 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1861 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1921 acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg
     1981 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca
     2041 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2101 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     2161 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2221 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca ggtaccggtg
     2281 aattcaccat ggacaaagac tgcgaaatga agcgcaccac cctggatagc cctctgggca
     2341 agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg ggcaaaggaa
     2401 catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc ggaccagagc
     2461 cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag gccatcgagg
     2521 agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt acccgccagg
     2581 tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac agccacctgg
     2641 ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg agcggaaatc
     2701 ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac gtggggggct
     2761 acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac agactgggca
     2821 agcctgggct gggtcctgca ggcggatccg cgtttaaact cgaggttaat taatgagcgg
     2881 ccgcatagat aactgatcca gtgtgctgga attaattcgc tgtctgcgag ggccagctgt
     2941 tggggtgagt actccctctc aaaagcgggc atgacttctg cgctaagatt gtcagtttcc
     3001 aaaaacgagg aggatttgat attcacctgg cccgcggtga tgcctttgag ggtggccgcg
     3061 tccatctggt cagaaaagac aatctttttg ttgtcaagct tgaggtgtgg caggcttgag
     3121 atctggccat acacttgagt gacaatgaca tccactttgc ctttctctcc acaggtgtcc
     3181 actcccaggt ccaactgcag gtcgagcatg catctagggc ggccaattcc gcccctctcc
     3241 cccccccccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3301 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3361 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3421 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3481 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3541 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3601 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3661 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     3721 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     3781 ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccgggata
     3841 attcctgcag ccaatatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
     3901 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3961 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4021 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4081 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4141 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4201 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4261 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4321 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4381 aggctcaagg cgcgcatgcc cgacggcgat gatctcgtcg tgacccatgg cgatgcctgc
     4441 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4501 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4561 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4621 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag gggatcaatt ctctagataa
     4681 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4741 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4801 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4861 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtcgag
     4921 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4981 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5041 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5101 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5161 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5221 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5281 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5341 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5401 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5461 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5521 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5581 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5641 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5701 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5761 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5821 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5881 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5941 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6001 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6061 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7141 catttccccg aaaagtgcca cctgacgtc
//
