LOCUS Exported 5260 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Contains human Histone H2B fused to EGFP in the pCS2+ expression vector. ACCESSION . VERSION . KEYWORDS pCS-H2B-EGFP 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" primer_bind complement(45..65) /label=T3 /note="T3 promoter, forward primer" promoter complement(45..63) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(84..100) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(84..100) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(97..119) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 108..124 /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(132..162) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 177..198 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(315..332) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(486..1074) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(566..585) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1245..2105) /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 1868..1887 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2106..2210) /gene="bla" /label=AmpR promoter rep_origin complement(2236..2691) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2373..2394) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2585..2604 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2765..3749 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 3784..3802 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3784..3801 /label=SP6 /note="SP6 promoter, forward primer" promoter complement(3871..3888) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3907..4284 /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSAK" regulatory 4297..4306 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4303..5022 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4303..5019 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4348..4369) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4609..4628) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4956..4977 /label=EGFP-C /note="EGFP, forward primer" promoter complement(5036..5053) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 5058..5192 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5082..5101) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5136..5155 /label=SV40pA-R /note="SV40 polyA, reverse primer" ORIGIN 1 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta 61 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 121 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 181 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 241 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 301 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 361 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 421 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 481 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 541 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 601 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 661 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 721 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 781 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 841 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 901 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 961 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 1021 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 1081 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 1141 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 1201 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 1261 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 1321 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 1381 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 1441 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 1501 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 1561 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 1621 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 1681 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 1741 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 1801 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 1861 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 1921 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 1981 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 2041 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 2101 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 2161 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 2221 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 2281 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa 2341 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa 2401 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac 2461 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga 2521 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa 2581 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc 2641 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc 2701 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 2761 agtcgaccat agccaattca atatggcgta tatggactca tgccaattca atatggtgga 2821 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga 2881 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga 2941 cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg 3001 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat 3061 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg 3121 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg 3181 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg 3241 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct 3301 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt 3361 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc 3421 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac 3481 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc 3541 aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat 3601 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg 3661 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg 3721 tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc 3781 ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga tcccatcgat 3841 tcgaattcaa ggcctctcga gcctctagaa ctatagtgag tcgtattact cgacggtacc 3901 gccaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg 3961 gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat 4021 tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag 4081 gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct 4141 tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc 4201 gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc 4261 atcaccaagt acaccagcgc taaggatcca ccggtcgcca ccatggtgag caagggcgag 4321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac 4381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag 4441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc 4501 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag 4561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac 4621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg 4681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac 4741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc 4801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac 4861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc 4921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc 4981 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccc tagaactata 5041 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca 5101 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat 5161 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt 5221 ttcaggttca gggggaggtg tgggaggttt tttaattcgc //