LOCUS       sequence-492433-        8091 bp    DNA     circular SYN 10-NOV-2025
DEFINITION  Expresses photoactivatable H2B-Cherry under CAG promoter and can be 
            randomly integrated by PiggyBac transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8091)
  AUTHORS   Nozaki T, Imai R, Tanbo M, Nagashima R, Tamura S, Tani T, Joti Y, 
            Tomita M, Hibino K, Kanemaki MT, Wendt KS, Okada Y, Nagai T, 
            Maeshima K
  TITLE     Dynamic Organization of Chromatin Domains Revealed by 
            Super-Resolution Live-Cell Imaging.
  JOURNAL   Mol Cell. 2017 Jul 20;67(2):282-293.e7. doi: 
            10.1016/j.molcel.2017.06.018. Epub 2017 Jul 14.
  PUBMED    28712725
REFERENCE   2  (bases 1 to 8091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2017.06.018"; journalName: "Mol Cell"; date: 
            "2017-07-20- 20"; volume: "67"; issue: "2"; pages: "282-293.e7"
COMMENT     Sequence Label: pPB-CAG-IB-H2B-PA-mCherry
FEATURES             Location/Qualifiers
     source          1..8091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(769..1044)
                     /label=chicken beta-actin promoter
     enhancer        1046..1425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   1711..1745
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1921..1940)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1922..1940)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1947..1964)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1947..1963)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1956..1978)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2105..2560
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2192..2211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2402..2423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2586..2690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2691..2759,2760..3551)
                     /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(2909..2928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3722..4310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4211..4230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4464..4481
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4598..4619
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4634..4639,4640..4657,4658..4664)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4672..4688
                     /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     4677..4699
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4696..4712
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4696..4712
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4733..4751
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   4881..4943
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     misc_feature    join(6058..6066,6067..6069,6070..6644)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6231..6250)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6460..6477
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(6672..7382)
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6950..6969)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7219..7237
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7379..7388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(7401..7778)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     primer_bind     complement(7816..7835)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          join(7843..8091,1..768)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7897..7919)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cctcccagat ttcggctccg cacagatttg ggacaaagga agtccctgcg ccctctcgca
       61 cgattaccat aaaaggcaat ggctgcggct cgccgcgcct cgacagccgc cggcgctccg
      121 ggggccgccg cgcccctccc ccgagccctc cccggcccga ggcggccccg ccccgcccgg
      181 cacccccacc tgccgccacc ccccgcccgg cacggcgagc cccgcgccac gccccgtacg
      241 gagccccgca cccgaagccg ggccgtgctc agcaactcgg ggaggggggt gcaggggggg
      301 ttgcagcccg accgacgcgc ccacaccccc tgctcacccc cccacgcaca caccccgcac
      361 gcagcctttg ttcccctcgc agcccccccc gcaccgcggg gcaccgcccc cggccgcgct
      421 cccctcgcgc acactgcgga gcgcacaaag ccccgcgccg cgcccgcagc gctcacagcc
      481 gccgggcagc gcggagccgc acgcggcgct ccccacgcac acacacacgc acgcaccccc
      541 cgagccgctc cccccgcaca aagggccctc ccggagcccc tcaaggcttt cacgcagcca
      601 cagaaaagaa acaagccgtc attaaaccaa gcgctaatta cagcccggag gagaagggcc
      661 gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag agccggggcg ggcggcgcga
      721 ggcggcggcg gagcggggca cggggcgaag gcagcgcgca gcgactcccg cccgccgcgc
      781 gcttcgcttt ttatagggcc gccgccgccg ccgcctcgcc ataaaaggaa actttcggag
      841 cgcgccgctc tgattggctg ccgccgcacc tctccgcctc gccccgcccc gcccctcgcc
      901 ccgccccgcc ccgcctggcg cgcscccccc ccccccccgc ccccatcgct gcacaaaata
      961 attaaaaaat aaataaatac aaaattgggg gtggggaggg gggggagatg gggagagtga
     1021 agcagaacgt ggggctcacc tcgaccatgg taatagcgat gactaatacg tagatgtact
     1081 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     1141 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     1201 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     1261 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     1321 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     1381 accccgtaat tgattactat taataactag tcaataatca atgtcgaccg ggatctgata
     1441 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa
     1501 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt
     1561 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct
     1621 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat
     1681 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc
     1741 tagggttaaa aaagatttgc gctttactcg acctaaactt taaacacgtc atagaatctt
     1801 cgtttgacaa aaaccacatt gtggccaagc tgtgtgacgc gacgcgcgct aaagaatggc
     1861 aaaccaagtc gcgcgaggta cctctcttaa ggtagcgcga tcgcacgcgt acccaattcg
     1921 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     1981 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2041 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2101 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2521 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2581 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2641 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2701 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2761 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2821 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2881 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2941 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3001 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3061 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3121 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3181 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3241 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3301 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3361 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3421 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3481 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3541 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3601 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3661 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3721 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3781 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3841 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3901 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3961 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4021 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4081 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4141 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4201 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4261 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4321 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4381 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4441 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4501 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4561 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4621 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4681 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc gaaattaacc
     4741 ctcactaaag ggaacaaaag ctggagctag ggataacagg gtaatggcgc gccttaatta
     4801 actataacgg tcctaaggta gcgagctcgg tattcacgac agcaggctga ataataaaaa
     4861 aattagaaac tattatttaa ccctagaaag ataatcatat tgtgacgtac gttaaagata
     4921 atcatgcgta aaattgacgc atgtgtttta tcggtctgta tatcgaggtt tatttattaa
     4981 tttgaataga tattaagttt tattatattt acacttacat actaataata aattcaacaa
     5041 acaatttatt tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa tttcttctat
     5101 aaagtaacaa agggcccgtt taaactcgac cagcttctga tggaattaga acttggcaaa
     5161 acaatactga gaatgaagtg tatgtggaac agaggctgct gatctcgttc ttcaggctat
     5221 gaaactgaca catttggaaa ccacagtact tagaaccaca aagtgggaat caagagaaaa
     5281 acaatgatcc cacgagagat ctatagatct atagatcatg agtgggagga atgagctggc
     5341 ccttaatttg gttttgcttg tttaaattat gatatccaac tatgaaacat tatcataaag
     5401 caatagtaaa gagccttcag taaagagcag gcatttatct aatcccaccc cacccccacc
     5461 cccgtagctc caatccttcc attcaaaatg taggtactct gttctcaccc ttcttaacaa
     5521 agtatgacag gaaaaacttc cattttagtg gacatcttta ttgtttaata gatcatcaat
     5581 ttctgcatcc cgtcaccatc aactcatgct cggttcatct agtaacagaa agtagcttga
     5641 atgtacaaga tcaatggctg gtgcttagcc ctcccacaca taaccagagg gcagcaattc
     5701 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     5761 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     5821 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     5881 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     5941 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     6001 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcatggt
     6061 tgtggccata ttatcatcgt gtttttcaaa ggaaaaccac gtccccgtgg ttcggggggc
     6121 ctagacgttt ttttaacctc gactaaacac atgtaaagca tgtgcaccga ggccccagat
     6181 cagatcccat acaatggggt accttctggg catccttcag ccccttgttg aatacgcttg
     6241 aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac tggggttgtg
     6301 ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac ctgtcgccag
     6361 gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt caagaagctt
     6421 ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg gcgagagggg
     6481 aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc ctcacattgc
     6541 caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg ccttattcca
     6601 agcggcttcg gccagtaacg ttaggggggg gggagggaga gggggggggg gcggaattcg
     6661 ttaacctcga gttacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggca
     6721 cgttcgtact gttccacgat ggtgtagtcc tcgttgtgtg aggtgatgtc caacttgcgg
     6781 ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc
     6841 tcagcgtcgt agtggccgcc gtccttcagc ttcactctcg gcttgacctc gcccttcagg
     6901 gcgccgtcct cggggtacat ccgctcggag agggcctccc agcccatggt cttcttctgc
     6961 attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactca
     7021 ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaatttcatc
     7081 acgcgctccc acttgaagcc ctcggggaag gacagcttaa agtagtcggg gatgtcggcg
     7141 gggtgcttca cgtaggcatt ggagccgtac atgaattgag gggacaggat gtcccaggtg
     7201 aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg
     7261 cggccctcgc cctcgccctc gatctcgaac acgtggccgt tcacggaccc ctccatgtgc
     7321 accttgaagc gcatgaactc cttaatgatg gccatgttat cctcctcgcc cttgctcacc
     7381 atggtggcga ccggtggatc cttagcgctg gtgtacttgg tgatggcctt agtaccctcg
     7441 gacacggcgt gcttggccaa ctccccaggc agcagcaggc gcacggccgt ctggatctcc
     7501 ctggaggtga tggtcgagcg cttgttgtaa tgcgccaggc gggaagcctc acctgcgatg
     7561 cgctcgaaaa tgtcgttcac aaacgaattc atgatgccca tggccttgga cgaaatgccg
     7621 gtgtcagggt ggacctgctt cagaaccttg tacacataga tggaatagct ctccttgcgg
     7681 ctgcgcttgc gcttcttgcc gcctttcttc tgcgccttag tcaccgcctt cttggagccc
     7741 tttttcgggg cgggagcaga cttcgctggc tctggcatct cgagagatca attctttgcc
     7801 aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga aaaagaagaa
     7861 ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga agccgaaccc
     7921 cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc gaggctggag
     7981 agggagaagg ggacggcggc gcggcgacgc acgaaggccc tccccgccca tttccttcct
     8041 gccggcgccg caccgcttcg ccccgcgccc gctagagggg gtgcggcggc g
//
