LOCUS Exported 4046 bp ds-DNA circular SYN 29-SEP-2022 DEFINITION Vector for generating dsDNA donors for homology-directed repair. Contains the visible marker 3xP3-DsRed.. ACCESSION . VERSION . KEYWORDS pHD-DsRed SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4046) AUTHORS Gratz SJ, Ukken FP, Rubinstein CD, Thiede G, Donohue LK, Cummings AM, O'Connor-Giles KM TITLE Highly specific and efficient CRISPR/Cas9-catalyzed homology-directed repair in Drosophila. JOURNAL Genetics. 2014 Apr;196(4):961-71. doi: 10.1534/genetics.113.160713. Epub 2014 Jan 29. PUBMED 24478335 REFERENCE 2 (bases 1 to 4046) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 29, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4046 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(106..139) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 365..374 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 371..1048 /codon_start=1 /product="rapidly maturing tetrameric variant of DsRed fluorescent protein (Bevis and Glick, 2002)" /label=DsRed-Express /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH LFL" primer_bind complement(501..519) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(551..571) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 769..788 /label=mCherry-F /note="mCherry, forward primer" primer_bind 963..986 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 1169..1290 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1206..1225) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1260..1279 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(1297..1330) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(1866..1883) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1866..1882) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1981..1998) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2152..2740) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2232..2251) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2862..3722) /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 3485..3504 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3723..3814) /gene="bla" /label=AmpR promoter primer_bind 3856..3875 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3881..3900 /label=pENTR-R /note="pENTR vectors, reverse primer" ORIGIN 1 aaagactggg cctttcgccc gggctaatta tggggtgtcg cccttcgctg aagcaggtgg 61 aattcttgca tgctagcggc cgcggacata tgcacacctg cgatcataac ttcgtataat 121 gtatgctata cgaagttatc gtacgggatc taattcaatt agagactaat tcaattagag 181 ctaattcaat taggatccaa gcttatcgat ttcgaaccct cgaccgccgg agtataaata 241 gaggcgcttc gtctacggag cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta 301 agcgaaagct aagcaaataa acaagcgcag ctgaacaagc taaacaatcg gctcgaagcc 361 ggtcgccacc atggcctcct ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg 421 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc 481 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc 541 ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc 601 cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat 661 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctcc aggacggctc 721 cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtaatgca 781 gaagaagact atgggctggg aggcgtccac cgagcgcctg tacccccgcg acggcgtgct 841 gaagggcgag atccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt 901 caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggactc 961 caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcgc 1021 cgagggccgc caccacctgt tcctgtaggg gccgcgactc tagatcataa tcagccatac 1081 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 1141 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 1201 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 1261 tggtttgtcc aaactcatca atgtatctta accggtataa cttcgtataa tgtatgctat 1321 acgaagttat agaagagcac tagtaaagat ctccatgcat aaggcgcgcc taggccttct 1381 gcagctcgag gctcttccgt caatcgagtt caagggcgac acaaaattta ttctaaatgc 1441 ataataaata ctgataacat cttatagttt gtattatatt ttgtattatc gttgacatgt 1501 ataattttga tatcaaaaac tgattttccc tttattattt tcgagattta ttttcttaat 1561 tctctttaac aaactagaaa tattgtatat acaaaaaatc ataaataata gatgaatagt 1621 ttaattatag gtgttcatca atcgaaaaag caacgtatct tatttaaagt gcgttgcttt 1681 tttctcattt ataaggttaa ataattctca tatatcaagc aaagtgacag gcgcccttaa 1741 atattctgac aaatgctctt tccctaaact ccccccataa aaaaacccgc cgaagcgggt 1801 ttttacgtta tttgcggatt aacgattact cgttatcaga accgcccagg gggcccgagc 1861 ttaagactgg ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa aaaggccatc 1921 cgtcaggggc cttctgctta gtttgatgcc tggcagttcc ctactctcgc cttccgcttc 1981 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 2041 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 2101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 2161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 2221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 2281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 2341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 2401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 2461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 2521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggg ctaactacgg 2581 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 2641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 2701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 2761 tacggggtct gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg gattttggtc 2821 atgagcttgc gccgtcccgt caagtcagcg taatgctctg cttaccaatg cttaatcagt 2881 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 2941 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg 3001 cgagaaccac gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc 3061 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 3121 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca 3181 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 3241 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 3301 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 3361 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 3421 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 3481 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 3541 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 3601 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 3661 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 3721 atattcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 3781 tacatatttg aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa 3841 ccaattctga acattatcgc gagcccattt atacctgaat atggctcata acaccccttg 3901 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa 3961 acgccgtagc gccgatggta gtgtggggac tccccatgcg agagtaggga actgccaggc 4021 atcaaataaa acgaaaggct cagtcg //