LOCUS       sequence-192969-        5575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein mRuby2
            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 5575)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5575)
  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            "Unpublished"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-mRuby2
FEATURES             Location/Qualifiers
     source          1..5575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..1002
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1037..1055
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1037..1054
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1106..1483
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1496..1505
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1508..1534
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1538..1564
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1568..1594
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2102..2128
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2135..2161
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2174..2200
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2207..2233
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2417..2443
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2447..2473
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2477..2503
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2504..2530
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2534..2560
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2564..2590
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2629..2763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2653..2672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2707..2726
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2876..2896)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2876..2894)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2915..2931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2915..2931)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2928..2950)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2939..2955
                     /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(2963..2969,2970..2987,2988..2993))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3008..3029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3146..3163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3317..3905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3397..3416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4076..4867,4868..4936))
                     /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     4699..4718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4937..5041)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5067..5522)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5204..5225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5416..5435
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcttcgctat tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat
       61 tcaatatggt ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat
      121 tcaatatggt ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat
      181 tcaatatggc ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa
      241 gtttgaggag gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa
      301 taatggcggc catattggct atatgccagg atcaatatat aggcaatatc caatatggcc
      361 ctatgccaat atggctattg gccaggttca atactatgta ttggccctat gccatatagt
      421 attccatata tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat
      481 accatatatg gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct
      541 ctatatatgg tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct
      601 cccccattga cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat
      661 aggaccaccc accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac
      721 gccccctatt gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta
      781 atagtaactt ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc
      841 cattgacgtc aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt
      901 caatggggag gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt
      961 gcctaatggg aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga
     1021 cgtcggagca agcttgattt aggtgacact atagaataca agctacttgt tctttttgca
     1081 ggatcccatc gacggtaccg ccaccatgcc agagccagcg aagtctgctc ccgccccgaa
     1141 aaagggctcc aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg
     1201 cagccgcaag gagagctatt ccatctatgt gtacaaggtt ctgaagcagg tccaccctga
     1261 caccggcatt tcgtccaagg ccatgggcat catgaattcg tttgtgaacg acattttcga
     1321 gcgcatcgca ggtgaggctt cccgcctggc gcattacaac aagcgctcga ccatcacctc
     1381 cagggagatc cagacggccg tgcgcctgct gctgcctggg gagttggcca agcacgccgt
     1441 gtccgagggt actaaggcca tcaccaagta caccagcgct aaggatccac cggtcgccac
     1501 catgggttat ccttacgatg tccctgatta cgctggctat ccttatgatg tcccggatta
     1561 tgctggctat ccgtatgatg ttcctgatta tgcaggtgga aattcgctga ttaaggagaa
     1621 tatgagaatg aaagttgtgc tcgaaggctc ggtgaatgga catcaattca aatgcactgg
     1681 agagggagaa ggtaacccgt acatgggaac acaaactatg cgaattaagg tgattgaggg
     1741 aggcccactt ccttttgctt ttgacatcct cgcaacaagt tttatgtacg gatcgagaac
     1801 atttatcaag tatccaaaag gaatcccaga ctttttcaaa caatcattcc cggaaggatt
     1861 tacttgggaa agagttacac gatatgaaga tggtggagtt atcaccgtga tgcaggatac
     1921 ttctctcgaa gatggatgtc tcgtttacca tgcacaagtt agaggagtca attttccatc
     1981 aaatggcgct gttatgcaga agaagacgaa gggatgggaa ccgaatactg agatgatgta
     2041 tccagctgat ggaggattac gcggatacac gcacatggct ttgaaagttg atggtggagg
     2101 atacccttat gatgtgccag actacgctgg aggttaccct tatgatgttc cagattatgc
     2161 tggcggagga ggataccctt atgacgttcc agattatgcc ggaggttacc cgtacgatgt
     2221 cccggactac gctggtggac acctttcctg ctcgttcgtc actacatata gatcgaaaaa
     2281 gaccgtggga aacatcaaga tgccaggaat ccatgctgtc gatcatagac tggaaagact
     2341 tgaagaatcc gataacgaga tgtttgttgt tcaacgcgag cacgcggtcg ctaaatttgc
     2401 aggacttggt ggtggatacc cgtatgacgt cccggattac gctggatacc cgtacgatgt
     2461 ccctgattat gcaggatacc cgtatgatgt gccggattat gcttacccat acgatgttcc
     2521 tgactatgcg ggctatccct atgacgtccc ggactatgca ggatatccat atgacgttcc
     2581 agattacgct ggcggatagt ctagaactat agtgagtcgt attacgtaga tccagacatg
     2641 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2701 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2761 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2821 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     2881 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2941 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     3001 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     3061 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     3121 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3181 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3241 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3481 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3721 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3841 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3901 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3961 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4021 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4081 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4141 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4201 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4261 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4321 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4381 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4441 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4501 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4561 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4621 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4681 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4741 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4801 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4861 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4921 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4981 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5041 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     5101 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5161 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5221 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5281 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5341 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5401 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5461 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5521 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcc
//
