LOCUS       sequence-68422-e        8842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Histone H2B fused with tandem iRFP, ires-puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8842)
  AUTHORS   Miyanari Y, Ziegler-Birling C, Torres-Padilla ME
  TITLE     Live visualization of chromatin dynamics with fluorescent TALEs.
  JOURNAL   Nat Struct Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680.
  PUBMED    24096363
REFERENCE   2  (bases 1 to 8842)
  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: "Nat Struct 
            Mol Biol. 2013 Oct 6. doi: 10.1038/nsmb.2680."
COMMENT     Sequence Label: pCAG-H2BtdiRFP-IP
FEATURES             Location/Qualifiers
     source          1..8842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          274..1290
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1214..1236
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1298..1317
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1395..1772
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1785..1794
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(3806..4380,4381..4383,4384..4392)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3973..3990)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4200..4219
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4393..4992
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4393..4412)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4889..4909
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5073..5090)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5079..5303
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5350..5366)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5350..5366)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5363..5385)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5374..5390
                     /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(5398..5404,5405..5422,5423..5428))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5443..5464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5523..5718
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5569..5704
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5724..5858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5926..5943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6097..6685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6177..6196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6856..7647,7648..7716))
                     /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     7479..7498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    8309..8443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        8457..8836
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8838..8842,1..273)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
       61 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
      121 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
      181 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      241 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
      301 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      361 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
      421 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
      481 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
      541 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
      601 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
      661 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
      721 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
      781 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
      841 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
      901 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
      961 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     1021 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     1081 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     1141 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     1201 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     1261 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     1321 tctcatcatt ttggcaaaga attatcacaa gtttgtacaa accccaggta cttgggaatt
     1381 cagatctcgc caccatgcca gagccagcga agtctgctcc cgccccgaaa aagggctcca
     1441 agaaggcggt gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg
     1501 agagctattc catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt
     1561 cgtccaaggc catgggcatc atgaattcgt ttgtgaacga cattttcgag cgcatcgcag
     1621 gtgaggcttc ccgcctggcg cattacaaca agcgctcgac catcacctcc agggagatcc
     1681 agacggccgt gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta
     1741 ctaaggccat caccaagtac accagcgcta aggatccacc ggtcgccacc atggccgaag
     1801 ggagcgtggc aagacagccc gatctgctga catgcgacga cgaaccaatc cacattccag
     1861 gagctatcca gcctcatgga ctgctgctgg ctctggccgc tgacatgact atcgtggcag
     1921 gctctgataa cctgccagag ctgaccgggc tggctatcgg agcactgatt ggccggtccg
     1981 cagccgacgt gttcgattct gagactcaca atagactgac catcgccctg gctgaacctg
     2041 gggctgcagt gggagcccca attaccgtcg gcttcacaat gcgaaaggac gctggcttta
     2101 tcgggtcctg gcaccgccat gatcagctga tttttctgga gctggaacca cctcagagag
     2161 acgtggcaga gccacaggct ttctttcgga gaacaaacag cgccatcagg cgcctgcagg
     2221 ccgctgagac tctggaatcc gcttgcgcag ccgctgcaca ggaagtgagg aagatcaccg
     2281 ggttcgatcg cgtcatgatc tacagattcg cctcagactt tagcggagaa gtgatcgcag
     2341 aagatcgctg tgccgaggtc gaaagtaaac tgggcctgca ttacccagcc tcaaccgtgc
     2401 ctgcacaggc acgacggctg tatacaatca accccgtgcg gatcattccc gacattaatt
     2461 acagaccagt gcccgtcaca cctgacctga acccagtgac tgggaggccc atcgatctga
     2521 gcttcgccat tctgcgcagc gtgtcccctg tccacctgga gtttatgagg aatatcggaa
     2581 tgcatggcac catgtctatc agtattctga gaggcgaaag gctgtggggg ctgatcgtgt
     2641 gccaccatcg gacaccttac tatgtggacc tggacggcag gcaggcttgt gaactggtgg
     2701 ctcaggtgct ggcttggcag attggcgtga tgaaggaggg gcatggcacc ggcagcaccg
     2761 gcagcggcag cagcggccgc tccggcaccg cctcctccga ggacaacaac atggccgccg
     2821 aagggagcgt ggcaagacag cccgatctgc tgacatgcga cgacgaacca atccacattc
     2881 caggagctat ccagcctcat ggactgctgc tggctctggc cgctgacatg actatcgtgg
     2941 caggctctga taacctgcca gagctgaccg ggctggctat cggagcactg attggccggt
     3001 ccgcagccga cgtgttcgat tctgagactc acaatagact gaccatcgcc ctggctgaac
     3061 ctggggctgc agtgggagcc ccaattaccg tcggcttcac aatgcgaaag gacgctggct
     3121 ttatcgggtc ctggcaccgc catgatcagc tgatttttct ggagctggaa ccacctcaga
     3181 gagacgtggc agagccacag gctttctttc ggagaacaaa cagcgccatc aggcgcctgc
     3241 aggccgctga gactctggaa tccgcttgcg cagccgctgc acaggaagtg aggaagatca
     3301 ccgggttcga tcgcgtcatg atctacagat tcgcctcaga ctttagcgga gaagtgatcg
     3361 cagaagatcg ctgtgccgag gtcgaaagta aactgggcct gcattaccca gcctcaaccg
     3421 tgcctgcaca ggcacgacgg ctgtatacaa tcaaccccgt gcggatcatt cccgacatta
     3481 attacagacc agtgcccgtc acacctgacc tgaacccagt gactgggagg cccatcgatc
     3541 tgagcttcgc cattctgcgc agcgtgtccc ctgtccacct ggagtttatg aggaatatcg
     3601 gaatgcatgg caccatgtct atcagtattc tgagaggcga aaggctgtgg gggctgatcg
     3661 tgtgccacca tcggacacct tactatgtgg acctggacgg caggcaggct tgtgaactgg
     3721 tggctcaggt gctggcttgg cagattggcg tgatgaagga ggaataactc gagagtgtac
     3781 aaagtggtga taattccgcc ccccccccct ctccctcccc cccccctaac gttactggcc
     3841 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     3901 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     3961 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     4021 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     4081 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     4141 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     4201 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     4261 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     4321 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     4381 atggccacaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4441 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4501 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4561 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4621 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4681 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4741 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4801 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4861 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4921 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4981 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     5041 catgcatcga tgatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     5101 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     5161 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     5221 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     5281 tggggatgcg gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc
     5341 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5401 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5461 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     5521 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5641 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5701 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     5761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5821 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     5881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6601 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6661 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6721 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6781 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6841 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6901 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6961 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7021 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7081 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7141 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7201 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7261 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7321 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7381 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7441 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7501 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7561 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7621 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7681 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatgaccgct
     7741 ttggaaaaac aaagactgta tttcctggaa attaatgttt attcaataaa ctgtgtattc
     7801 agctatattc acatagtggt gaggctgaaa tgaggcggga agaggcggtt ggggcttaat
     7861 tatatcaatt tgggtggccc cacagcgcct ccaaggcgcc agtcctgttt tgacaagttg
     7921 cctctggaag cctctcttct ttttctccag agtaagcgga ggccaggggc ccccggcctc
     7981 tgcttaatac taaaaaaaac agctgttgtc atagtaatga ttgggtggaa acattccagg
     8041 cctgggtgga gaggcttttt gcttcctctt gcaaaaccac actgacattc caggcctggg
     8101 tggagaggct ttttgcttcc tcttgcaaaa ccacactgcc ctctggaggg cagttgccta
     8161 gcaactaatt aaaagaggat gtcgcacggc cagctgcggt cagttagtca cttcctgctt
     8221 aactgacttg acattttcta ttttaagagt cgggaggaaa attactgtgt tggaggccct
     8281 ccgccatctt ctgaagctga atcgaattaa cttgtttatt gcagcttata atggttacaa
     8341 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8401 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgatatc atcgtcgaca
     8461 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     8521 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     8581 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     8641 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8701 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8761 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     8821 catcgctatt accatggtcg ag
//
