LOCUS       sequence-364585-        6496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of 6xHis CBX5 (HP1a) mCherry Fusion Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  AUTHORS   Asimi V, Sampath Kumar A, Niskanen H, Riemenschneider C, Hetzel S, 
            Naderi J, Fasching N, Popitsch N, Du M, Kretzmer H, Smith ZD, 
            Weigert R, Walther M, Mamde S, Meierhofer D, Wittler L, Buschow R, 
            Timmermann B, Cisse II, Ameres SL, Meissner A, Hnisz D
  TITLE     Hijacking of transcriptional condensates by endogenous retroviruses.
  JOURNAL   Nat Genet. 2022 Aug;54(8):1238-1247. doi: 
            10.1038/s41588-022-01132-w. Epub 2022 Jul 21.
  PUBMED    35864192
REFERENCE   2  (bases 1 to 6496)
  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"; doi: 
            "10.1038/s41588-022-01132-w"; journalName: "Nat Genet"; date: 
            "2022-08"; volume: "54"; issue: "8"; pages: "1238-1247"
COMMENT     Sequence Label: pET-45b-HP1a-mCherry
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(134..589)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(271..292)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     483..502
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(626..673)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     669..687
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(725..769)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(2093..2113)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2123..2140)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2150..2172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2187..2211
                     /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)."
     primer_bind     complement(2211..2230)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2212..2230)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2318..2337)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2543..2620
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2621..3703
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2640..2659)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4275..4466
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4460..4482)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4568..4707
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4623..4642
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4722..4739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4893..5481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4973..4992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5562..5654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5655..5723,5724..6496,1..19)
                     /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     complement(5873..5892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc
      121 atcgtgccgc cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
      181 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
      241 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      301 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      361 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      421 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      481 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      541 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
      601 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
      661 ggggttatgc tagttattgc tcagcggtgg cagcagccta ggttaattaa gctgcgctag
      721 tagacgagtc catgtgctgg cgttcaaatt tcgcagcagc ggtttcttta ccagactcga
      781 gtgcggccgc acagctcttc gcgctttctt tttctttgtt ttccgcatcc tctggatatg
      841 cgtgccacgt cagtctctct tcataaaatg ctatcacaat ctgtggacac ttcacgttag
      901 cttcttttgc aagaaccagg tcagcttcat ctgtgtcttt ccatttcatt aagaacatta
      961 agtcaccgca ggaatctgtt gctccgatga tcttttctgg ttccagtcct ctctcaaagc
     1021 cccgagcgat atcattgctt tgctctctct tttttttaga tttaatatca tcagcgctgt
     1081 tggagaaact ggatttcctc ttgtttcctt ctgatttctc cctgggctta ttgttttcac
     1141 cctccttcat cttcttatac tttttcataa actcagaaat tagttcagga caatccaagt
     1201 tcttctcagg ttcccaagta ttgtgctcct cagaaaagcc tttccacttc aacagatatt
     1261 ccacttgccc cttaaccatg cgcctgtcca acaccttttc caccacatat tcctcctcat
     1321 cctctgaaga agagctgtcg gctgtcctct tggtcttctt tcccatatcc gtgtacttgt
     1381 acaattcatc catgccccct gtgctatgtc tgccctcggc ccgctcgtac tgttccacga
     1441 tggtatagtc ttcgttatgg gatgtaatat ccagtttgat attcacgttg tatgccccag
     1501 gaagctgaac gggtttcttg gccttatagg tggttttgac ctcagcatca taatgtcccc
     1561 catccttcag cttgagtcgc tgcttgattt ctcccttcag tgctccgtcc tcgggataca
     1621 tgcgttctga gctggcttcc cagcccatag ttttcttctg catcacgggg ccgtctgatg
     1681 ggaagttagt gccccgaagt ttaaccttat agataaattc tccgtcctga agagagctgt
     1741 cctgggtaac tgtgactaca ccgccatcct caaaattcat tacccgttcc cacttgaacc
     1801 cttcagggaa gctcagtttc aagtagtccg gaatatcggc agggtgctta acgtaagcct
     1861 tagagccgta cataaactgg ggtgacagga tgtcccaggc gaaaggcagg gggccaccct
     1921 tggtcacctt cagtttggcc gtctgtgtcc cttcataggg tcgcccctcg ccttcgccct
     1981 cgatttcgaa ctcgtgaccg ttcacagatc cctccatatg caccttgaac cgcataaact
     2041 ctttaattat agccatatta tcctcctctc ctttggaaac cattccagaa ccggattgga
     2101 agtacaggtt ttcctcgatc ccatggtgat ggtgatggtg catggtatat ctccttctta
     2161 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
     2221 agtcgtatta atttcgcggg atcgagatcg atctcgatcc tctacgccgg acgcatcgtg
     2281 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     2341 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     2401 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     2461 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     2521 aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc gcggtatggc
     2581 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     2641 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     2701 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     2761 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     2821 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     2881 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     2941 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     3001 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     3061 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     3121 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     3181 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     3241 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     3301 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     3361 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     3421 catctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     3481 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     3541 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     3601 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3661 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     3721 taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc
     3781 cagtcagctc cttccggtgg gcgcggggca tgactagcat gatcgtgctc ctgtcgttga
     3841 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     3901 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     3961 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     4021 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     4081 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     4141 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     4201 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     4261 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     4321 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     4381 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     4441 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     4501 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     4561 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     4621 tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga
     4681 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     4741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5521 tggaacgaaa actcacgtta agggattttg gtcatgacct atttgtttat ttttctaaat
     5581 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aatcatgatt
     5641 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5701 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5761 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5821 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5881 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5941 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6001 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6061 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6121 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6181 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6241 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6301 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6361 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6421 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6481 ctgagatagg tgcctc
//
