LOCUS       sequence-192951-        5281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            Katushka2 in the pCS2+ expression vector. It has the SP6 RNA 
            polymerase site for in vitro transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5281)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5281)
  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            "Unpublished"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: pCS2+ H2B-Katushka2
FEATURES             Location/Qualifiers
     source          1..5281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..997
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1032..1050
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1032..1049
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1101..1478
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1491..1500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2205..2231
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2235..2261
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2265..2291
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2330..2464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2354..2373)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2408..2427
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2577..2597)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2577..2595)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2616..2632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2616..2632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2629..2651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2640..2656
                     /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(2664..2670,2671..2688,2689..2694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2709..2730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2847..2864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3018..3606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3098..3117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3777..4568,4569..4637))
                     /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     4400..4419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4638..4742)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4768..5223)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4905..4926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5117..5136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg ccaattcaat
       61 atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg ccaattcaat
      121 atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg ccaattcaat
      181 atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt gccaagtttg
      241 aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt gccaataatg
      301 gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata tggccctatg
      361 ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat atagtattcc
      421 atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc catataccat
      481 atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat ggtctctata
      541 tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg cgcctccccc
      601 attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg tcaataggac
      661 cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt ctcacgcccc
      721 ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc tattaatagt
      781 aacttggcaa gtacattact attggaagta cgccagggta cattggcagt actcccattg
      841 acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt gacgtcaatg
      901 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
      961 atgggaggtc tatataagca atgctcgttt agggaaccgc cattctgcct ggggacgtcg
     1021 gagcaagctt gatttaggtg acactataga atacaagcta cttgttcttt ttgcaggatc
     1081 ccatcgacgg taccgccacc atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg
     1141 gctccaagaa ggcggtgact aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc
     1201 gcaaggagag ctattccatc tatgtgtaca aggttctgaa gcaggtccac cctgacaccg
     1261 gcatttcgtc caaggccatg ggcatcatga attcgtttgt gaacgacatt ttcgagcgca
     1321 tcgcaggtga ggcttcccgc ctggcgcatt acaacaagcg ctcgaccatc acctccaggg
     1381 agatccagac ggccgtgcgc ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg
     1441 agggtactaa ggccatcacc aagtacacca gcgctaagga tccaccggtc gccaccatgg
     1501 tatccgtgct cataaccgaa aatatgcaca tgaagctgta catggaagga acagtcaacg
     1561 atcaccattt caagtgtacc agtgagggag agggcaaacc ctatgaggga acccagacca
     1621 tgaagatcaa ggccgttgaa ggtggaccct tgcctttcgc ctttgacatt ctggccacta
     1681 gctttatgta cggaagtaaa acctttatca atcacacgca aggcatccct gatttcttca
     1741 agcagagctt tccagagggc ttcacctggg agaggattac cacctacgag gacggaggcg
     1801 ttctgacggc cactcaggat acgtccttgc agaacggttg cctgatttac aatgttaaga
     1861 tcaacggcgt caactttcca tccaacggtc cggtaatgca gaagaagact ctgggctggg
     1921 aggccagcac cgaaaccctt taccccgctg acagcggact gcgcggtcac agccagatgg
     1981 ctctgaagtt ggtgggtggt ggatacctgc attgcagcct caagaccacc taccgctcca
     2041 agaaacccgc caaaaatctc aagatgcccg gcttctactt tgtagatcga cggttggagc
     2101 ggatcaagga ggcggataag gagacgtatg ttgagcagca tgagatggcg gtggcacggt
     2161 attgcgattt gccgtcgaaa ctgggtggta ccagcggcac cgtttaccca tacgatgttc
     2221 ctgactatgc gggctatccc tatgacgtcc cggactatgc aggatatcca tatgacgttc
     2281 cagattacgc tggcggatag tctagaacta tagtgagtcg tattacgtag atccagacat
     2341 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2401 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2461 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2521 tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt
     2581 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2641 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     2701 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2761 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2821 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2881 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2941 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3001 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3061 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3121 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3181 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3241 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3301 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3361 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3421 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3481 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3541 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3601 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3661 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3721 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3901 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4741 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     4801 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4861 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4921 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4981 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5041 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5101 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5161 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5221 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5281 c
//
