LOCUS       sequence-83995-e        5259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains fusion of human Histone H2B to citrine. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5259)
  AUTHORS   Megason SG
  TITLE     In toto imaging of embryogenesis with confocal time-lapse 
            microscopy.
  JOURNAL   Methods Mol Biol. 2009;546:317-32. doi: 
            10.1007/978-1-60327-977-2_19.
  PUBMED    19378112
REFERENCE   2  (bases 1 to 5259)
  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: "Methods Mol
            Biol."; date: "2009"; volume: "546:317-32. doi"; pages: " 
            10.1007/978-1-60327-977-2_19"
COMMENT     Sequence Label: pCS-H2B-citrine
FEATURES             Location/Qualifiers
     source          1..5259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        71..1055
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1090..1108
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1090..1107
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1206..1583
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1602..1611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1608..1610,1611..1613,1614..2324)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1653..1674)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1914..1933)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2261..2282
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2363..2497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2387..2406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2441..2460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2610..2630)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2610..2628)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2649..2665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2649..2665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2662..2684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2673..2689
                     /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(2697..2703,2704..2721,2722..2727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2742..2763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2880..2897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3051..3639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3131..3150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3810..4601,4602..4670))
                     /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     4433..4452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4671..4775)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4801..5256)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4938..4959)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5150..5169
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
       61 tacgccagtc gaccatagcc aattcaatat ggcgtatatg gactcatgcc aattcaatat
      121 ggtggatctg gacctgtgcc aattcaatat ggcgtatatg gactcgtgcc aattcaatat
      181 ggtggatctg gaccccagcc aattcaatat ggcggacttg gcaccatgcc aattcaatat
      241 ggcggacttg gcactgtgcc aactggggag gggtctactt ggcacggtgc caagtttgag
      301 gaggggtctt ggccctgtgc caagtccgcc atattgaatt ggcatggtgc caataatggc
      361 ggccatattg gctatatgcc aggatcaata tataggcaat atccaatatg gccctatgcc
      421 aatatggcta ttggccaggt tcaatactat gtattggccc tatgccatat agtattccat
      481 atatgggttt tcctattgac gtagatagcc cctcccaatg ggcggtccca tataccatat
      541 atggggcttc ctaataccgc ccatagccac tcccccattg acgtcaatgg tctctatata
      601 tggtctttcc tattgacgtc atatgggcgg tcctattgac gtatatggcg cctcccccat
      661 tgacgtcaat tacggtaaat ggcccgcctg gctcaatgcc cattgacgtc aataggacca
      721 cccaccattg acgtcaatgg gatggctcat tgcccattca tatccgttct cacgccccct
      781 attgacgtca atgacggtaa atggcccact tggcagtaca tcaatatcta ttaatagtaa
      841 cttggcaagt acattactat tggaaggacg ccagggtaca ttggcagtac tcccattgac
      901 gtcaatggcg gtaaatggcc cgcgatggct gccaagtaca tccccattga cgtcaatggg
      961 gaggggcaat gacgcaaatg ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat
     1021 gggaggtcta tataagcaat gctcgtttag ggaaccgcca ttctgcctgg ggacgtcgga
     1081 gcaagcttga tttaggtgac actatagaat acaagctact tgttcttttt gcaggatccc
     1141 atcgattcga attcaaggcc tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg
     1201 ccaccatgcc agagccagcg aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg
     1261 tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg cagccgcaag gagagctatt
     1321 ccatctatgt gtacaaggtt ctgaagcagg tccaccctga caccggcatt tcgtccaagg
     1381 ccatgggcat catgaattcg tttgtgaacg acattttcga gcgcatcgca ggtgaggctt
     1441 cccgcctggc gcattacaac aagcgctcga ccatcacctc cagggagatc cagacggccg
     1501 tgcgcctgct gctgcctggg gagttggcca agcacgccgt gtccgagggt actaaggcca
     1561 tcaccaagta caccagcgct aaggatcccc gggtaccggt cgccaccatg gtgagcaagg
     1621 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1681 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1741 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1801 tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1861 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1921 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1981 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2041 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2101 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2161 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2221 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2281 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccctagaa
     2341 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     2401 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2461 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2521 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     2581 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     2641 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2701 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2761 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2821 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2881 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2941 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3481 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3721 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3781 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3841 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3901 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3961 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4021 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4081 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4141 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4201 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4261 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4321 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4381 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4441 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4501 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4561 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4621 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4681 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4741 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4801 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4861 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4921 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4981 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5041 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5101 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5161 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5221 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccc
//
