LOCUS Exported 4048 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 4048) 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 4048) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 29, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4048 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 622..641 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(860..951) /gene="bla" /label=AmpR promoter primer_bind 993..1012 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1018..1037 /label=pENTR-R /note="pENTR vectors, reverse primer" protein_bind complement(1289..1322) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 1548..1557 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1554..2231 /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(1684..1702) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(1734..1754) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 1952..1971 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2146..2169 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 2352..2473 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2389..2408) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2443..2462 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(2480..2513) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(3049..3066) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3049..3065) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3164..3181) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3335..3923) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3415..3434) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4047..859) /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" ORIGIN 1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc 181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt 901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca 961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg 1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg 1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga 1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg 1201 cccgggctaa ttatggggtg tcgcccttcg ctgaagcagg tggaattctt gcatgctagc 1261 ggccgcggac atatgcacac ctgcgatcat aacttcgtat aatgtatgct atacgaagtt 1321 atcgtacggg atctaattca attagagact aattcaatta gagctaattc aattaggatc 1381 caagcttatc gatttcgaac cctcgaccgc cggagtataa atagaggcgc ttcgtctacg 1441 gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa 1501 taaacaagcg cagctgaaca agctaaacaa tcggctcgaa gccggtcgcc accatggcct 1561 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga 1621 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 1681 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc 1741 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca 1801 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 1861 gcgtggtgac cgtgacccag gactcctccc tccaggacgg ctccttcatc tacaaggtga 1921 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct 1981 gggaggcgtc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca 2041 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg 2101 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct 2161 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc 2221 tgttcctgta ggggccgcga ctctagatca taatcagcca taccacattt gtagaggttt 2281 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa 2341 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 2401 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 2461 tcaatgtatc ttaaccggta taacttcgta taatgtatgc tatacgaagt tatagaagag 2521 cactagtaaa gatctccatg cataaggcgc gcctaggcct tctgcagctc gaggctcttc 2581 cgtcaatcga gttcaagggc gacacaaaat ttattctaaa tgcataataa atactgataa 2641 catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt tgatatcaaa 2701 aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt aacaaactag 2761 aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta taggtgttca 2821 tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca tttataaggt 2881 taaataattc tcatatatca agcaaagtga caggcgccct taaatattct gacaaatgct 2941 ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg 3001 attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac tggccgtcgt 3061 tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc 3121 ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc actgactcgc 3181 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 3241 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 3301 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 3361 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 3421 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 3481 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 3541 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 3601 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 3661 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 3721 taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact agaagaacag 3781 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 3841 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 3901 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 3961 agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagct tgcgccgtcc 4021 cgtcaagtca gcgtaatgct ctgctttt //