LOCUS       sequence-242469-        5186 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to monomeric red fluorescent 
            protein 1 in the pCS2+ expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5186)
  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 5186)
  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-mRFP1
FEATURES             Location/Qualifiers
     source          1..5186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..1034
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1069..1087
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1069..1086
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1192..1569
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1582..1591
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1588..2265
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1588..2265
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1718..1736)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1986..2005
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2180..2203
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2272..2406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2296..2315)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2350..2369
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2519..2539)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2519..2537)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2558..2574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2571..2593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2582..2598
                     /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(2606..2612,2613..2630,2631..2636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2651..2672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2789..2806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2960..3548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3040..3059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3719..4510,4511..4579))
                     /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     4342..4361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4580..4684)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4710..5165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4847..4868)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5059..5078
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca
       61 attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca
      121 attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca
      181 attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca
      241 actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc
      301 aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca
      361 ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt
      421 caatactatg tattggccct atgccatata gtattccata tatgggtttt cctattgacg
      481 tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc
      541 catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca
      601 tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg
      661 gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg
      721 atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa
      781 tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt
      841 ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc
      901 gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg
      961 gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg
     1021 ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca
     1081 ctatagaata caagctactt gttctttttg caggatccca tcgattcgaa ttcaaggcct
     1141 ctcgagcctc tagaactata gtgagtcgta ttactcgacg gtaccgccac catgccagag
     1201 ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga aggcggtgac taaggcgcag
     1261 aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga gctattccat ctatgtgtac
     1321 aaggttctga agcaggtcca ccctgacacc ggcatttcgt ccaaggccat gggcatcatg
     1381 aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg aggcttcccg cctggcgcat
     1441 tacaacaagc gctcgaccat cacctccagg gagatccaga cggccgtgcg cctgctgctg
     1501 cctggggagt tggccaagca cgccgtgtcc gagggtacta aggccatcac caagtacacc
     1561 agcgctaagg atccaccggt cgccaccatg gcctcctccg aggacgtcat caaggagttc
     1621 atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1681 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     1741 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg ctccaaggcc
     1801 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     1861 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1921 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1981 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcggatgtac
     2041 cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc
     2101 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
     2161 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
     2221 gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaatacgt agatccagac
     2281 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2341 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2401 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2461 gttttttaat tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc
     2521 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2581 attgttatcc gctcacaatt ccacacaaca tacgagccgg gagcataaag tgtaaagcct
     2641 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2701 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2761 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4681 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     4741 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4801 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4861 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4921 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4981 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5041 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5101 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5161 cgcgtcccat tcgccattca ggctgc
//
