LOCUS       sequence-181910-        1779 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            mCerulean in the pCS2+ expression vector. It has the SP6 RNA 
            polymerase site for in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1779)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1779)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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            "Unpublished"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-mCerulean
FEATURES             Location/Qualifiers
     source          1..1779
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        94..112
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     94..111
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             163..540
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      553..562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(559..561,562..564,565..1275)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(604..625)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(865..884)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1212..1233
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1279..1344
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1387..1521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1411..1430)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1465..1484
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1634..1654)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1634..1652)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1673..1689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1673..1689)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1686..1708)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1697..1713
                     /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(1721..1727,1728..1745,1746..1751))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaacc
       61 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
      121 tacttgttct ttttgcagga tcccatcgac ggtaccgcca ccatgccaga gccagcgaag
      181 tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc
      241 ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg
      301 aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt
      361 gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag
      421 cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag
      481 ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag
      541 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      601 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      661 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      721 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc
      781 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      841 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      901 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      961 ggcaacatcc tggggcacaa gctggagtac aacgccatca gcgacaacgt ctatatcacc
     1021 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     1081 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1141 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     1201 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1261 gacgagctgt acaagatgga ctacaaagac catgacggtg attataaaga tcatgacatc
     1321 gattacaagg atgacgatga caagtaacgt aagctagtct agaactatag tgagtcgtat
     1381 tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1441 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1501 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1561 ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac
     1621 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     1681 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     1741 taaagtgtaa agcctggggt gcctaatgag tgagctaac
//
