LOCUS       sequence-83988-e        5260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to cerulean fluorescent 
            protein in the pCS2+ vector. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5260)
  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 5260)
  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-cerulean
FEATURES             Location/Qualifiers
     source          1..5260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(13..15,16..18,19..732)
                     /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(58..79)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(319..338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     666..687
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    768..902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(792..811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     846..865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1015..1035)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1015..1033)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1054..1070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1054..1070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1067..1089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1078..1094
                     /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(1102..1108,1109..1126,1127..1132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1147..1168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1285..1302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1456..2044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1536..1555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2215..3006,3007..3075))
                     /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     2838..2857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3076..3180)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3206..3661)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3343..3364)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3555..3574
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3735..4719
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        4754..4772
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4754..4771
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4877..5254
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
ORIGIN
        1 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
       61 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      121 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      181 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcgc ccgctacccc
      241 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      301 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      361 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      421 atcctggggc acaagctgga gtacaacgcc atcagcgaca acgtctatat caccgccgac
      481 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
      541 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      601 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
      661 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      721 ctgtacaagt aaagcggccc tagaactata gtgagtcgta ttacgtagat ccagacatga
      781 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      841 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      901 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      961 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     1021 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1081 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1141 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1201 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1261 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1321 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1861 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     1921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2101 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2161 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2221 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2281 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2341 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2401 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2461 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2521 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2581 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2641 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2701 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2761 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2821 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2881 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2941 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3001 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3061 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3121 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3181 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     3241 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3301 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3361 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3421 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3481 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3541 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3601 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3661 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     3721 ctattacgcc agtcgaccat agccaattca atatggcgta tatggactca tgccaattca
     3781 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     3841 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     3901 atatggcgga cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     3961 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     4021 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     4081 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     4141 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     4201 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     4261 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     4321 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     4381 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     4441 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     4501 gtaacttggc aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat
     4561 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     4621 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     4681 taatgggagg tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt
     4741 cggagcaagc ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga
     4801 tcccatcgat tcgaattcaa ggcctctcga gcctctagaa ctatagtgag tcgtattact
     4861 cgacggtacc gccaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc
     4921 caagaaggcg gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa
     4981 ggagagctat tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat
     5041 ttcgtccaag gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc
     5101 aggtgaggct tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat
     5161 ccagacggcc gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg
     5221 tactaaggcc atcaccaagt acaccagcgc taaggatcca
//
