LOCUS       sequence-209767-        5403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human HIST1H2BJ.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5403)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5403)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-52: HIST1H2BJ-mEGFP
FEATURES             Location/Qualifiers
     source          1..5403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     194..211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     195..211
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             854..1231
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             join(1250..1252,1253..1966)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1292..1313)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1553..1572)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1900..1921
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2998..3014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2998..3014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3011..3033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3022..3038
                     /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(3046..3052,3053..3070,3071..3076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3091..3112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3229..3246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3400..3988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3480..3499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4159..4950,4951..5019))
                     /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     4782..4801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5020..5124)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5192..5210
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5248..5270)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5370..5389
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
       61 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      181 ccagtcacga cgttgtaaaa cgacggccag tgaattgacg cgtattggga tctttgccca
      241 gcagcttgtt gagctcctcg tcgttgcgga tggccagttg gagatgacgc gggatgatgc
      301 gggtcttctt gttgtcgcgg gccgcgttgc ccgccagttc caggatctcg gcggtcaggt
      361 actccagcac cgctgccaga tacaccggcg cgccggcccc gacccgctcg gcatagttgc
      421 ctttgcggag caggcggtgc actcggccca cggggaactg gagaccggcc ctagaagagc
      481 gagtcttggc cttagcgcgg gctttgcctc cctgcttgcc acgtccagac atagcgagcg
      541 caactcacta cgagcaacca caaagtgaac gggaaaggcg gcgcttttta taaacactat
      601 tgggcgcgaa aaagaagacg tgttgttggt tagggctgca gtttaatttc aaccaatagt
      661 agtgcgtctt ctggatttgc gaatcctgat tgggcagacc tgacctctga cgttaccctg
      721 aataactacc aatcagacac aagacttcaa ctcttcacct tatttgcata agcgattcta
      781 tataaaagcg ccttgtcata ccctgctcac gctgtttttc cttttcgttg gcgctttata
      841 gctacacagt gctatgccag agccagcgaa gtctgctccc gccccgaaaa agggctccaa
      901 gaaggcggtg actaaggcgc agaagaaaga cggcaagaag cgcaagcgca gccgcaagga
      961 gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca ccggcatttc
     1021 gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg
     1081 tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca gggagatcca
     1141 gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt ccgagggtac
     1201 taaggccgtc accaagtaca ccagcgctaa ggaccctcct gtggccaccg tgagcaaggg
     1261 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1321 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1381 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1441 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1501 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1561 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1621 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1681 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1741 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1801 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1861 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1921 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaacag tgagttggtt
     1981 gcaaactctc aatcctaacg cctcttttaa gagccaccca tgttctcaaa gaaagagctg
     2041 gtgcttgtat tcctcctctg ctggccactg acaaaccctt gtaacttgct actgtgtttt
     2101 ttggtctgaa gtagagcagt tatttaacta atccttagtg actttttttt tttagatctg
     2161 ccattctaat cttagagtta agtaaggaga tgggaaattt tctattataa gttcgaaacc
     2221 aattaaaata cgttagaaac caattaaaat actcgtcggt cccccgtcgg ttagtgattt
     2281 ggaacagtgc caagttgcag cggttgtcag tttgaatttg cccgggcaac gcccgccctt
     2341 cctctgcatc ctgattggtt gttgtatcca ggaagtccac ggtcggcttg ggtgcgccaa
     2401 aggaatccaa accccgcctt tatttggcct tccttacaaa ctcagtaaac tgtcagtttc
     2461 tcattcttta gtattttatc gctttcggga aggagattta tttctaatta gttttcgttg
     2521 gatagagcgt tttttttttt aaagggaccc tttatggcaa cataatcagt tttttttttt
     2581 ttgagaagga gacttcctct gtcgcccaga ctggagtgcg gtggcgcgat ctcagctcac
     2641 tgcagcctcc gcctcccggg ttccagcgat tctcctgctt cacccttccg tgtagctggg
     2701 attacaggca tgcgccacca tgcccggata atttttgtat ttttagtaga gatggggttt
     2761 caccatgttg gctaggctgg tctcgaactg acttcaggtg atccactagc ctcagcctta
     2821 aagtgctagg attacaggca tgagccatca agcccggcta caacggaatt tgaatgctgt
     2881 agttttcagc ctgagaaaga tcaatgcttt atttctgatt agaatactat tttcccttta
     2941 tggtatatct atataagcaa aggatcccaa tggcgcgccg agcttggcgt aatcatggtc
     3001 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3061 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3121 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3181 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3241 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3361 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4081 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4141 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4201 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4261 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4321 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4381 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4441 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4501 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4561 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4621 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4681 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4741 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4801 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4861 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4921 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4981 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5041 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5101 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5161 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5221 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5281 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5341 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5401 cca
//
