LOCUS Exported 5260 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Contains a fusion of human histone H2B to cerulean fluorescent protein in the pCS2+ vector. . ACCESSION . VERSION . KEYWORDS pCS-H2B-cerulean 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5260 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 7..16 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 13..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" promoter complement(746..763) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 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(1102..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(2215..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" promoter complement(4841..4858) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 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 //