LOCUS       sequence-222558-        4096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating dsDNA donors for homology-directed repair to 
            replace genes or other genomic sequence with an attP docking site. 
            Contains the visible marker 3xP3-DsRed. As known as pHD-DsRed-attP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4096)
  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 4096)
  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"; doi: 
            "10.1534/genetics.113.160713"; journalName: "Genetics"; date: 
            "2014-04"; volume: "196"; issue: "4"; pages: "961-71"
COMMENT     Sequence Label: pDsRed-attP
FEATURES             Location/Qualifiers
     source          1..4096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(257..1048,1049..1117))
                     /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     880..899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1118..1209)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1251..1270
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1276..1295
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1597..1630
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1856..1865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1862..2539
                     /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(1992..2010)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2042..2062)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2260..2279
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2454..2477
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2660..2781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2697..2716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2751..2770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2788..2821
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3357..3374)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3357..3373)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3643..4096,1..135))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3723..3742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
       61 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      121 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      181 gacgcgcgcg taactcacgt taagggattt tggtcatgag cttgcgccgt cccgtcaagt
      241 cagcgtaatg ctctgcttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      301 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      361 gcttaccatc tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg
      421 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      481 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      541 ttaatagttt gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt
      601 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      661 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      721 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      781 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      841 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
      901 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      961 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1021 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1081 agggaataag ggcgacacgg aaatgttgaa tactcatatt cttccttttt caatattatt
     1141 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1201 ataaacaaat aggggtcagt gttacaacca attaaccaat tctgaacatt atcgcgagcc
     1261 catttatacc tgaatatggc tcataacacc ccttgtttgc ctggcggcag tagcgcggtg
     1321 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     1381 gggactcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     1441 gaaagactgg gcctttcgcc cgggctaatt atggggtgtc gcccttcgct gaagcaggtg
     1501 gaattcttgc atgctagcgg ccgcggacat atgcacacct gcgatcgtag tgccccaact
     1561 ggggtaacct ttgagttctc tcagttgggg gcgtagataa cttcgtataa tgtatgctat
     1621 acgaagttat cgtacgggat ctaattcaat tagagactaa ttcaattaga gctaattcaa
     1681 ttaggatcca agcttatcga tttcgaaccc tcgaccgccg gagtataaat agaggcgctt
     1741 cgtctacgga gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc
     1801 taagcaaata aacaagcgca gctgaacaag ctaaacaatc ggctcgaagc cggtcgccac
     1861 catggcctcc tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg
     1921 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     1981 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     2041 cctgtccccc cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc
     2101 cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     2161 ggacggcggc gtggtgaccg tgacccagga ctcctccctc caggacggct ccttcatcta
     2221 caaggtgaag ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac
     2281 tatgggctgg gaggcgtcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
     2341 gatccacaag gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtccat
     2401 ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggact ccaagctgga
     2461 catcacctcc cacaacgagg actacaccat cgtggagcag tacgagcgcg ccgagggccg
     2521 ccaccacctg ttcctgtagg ggccgcgact ctagatcata atcagccata ccacatttgt
     2581 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2641 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2761 caaactcatc aatgtatctt aaccggtata acttcgtata atgtatgcta tacgaagtta
     2821 tagaagagca ctagtaaaga tctccatgca taaggcgcgc ctaggccttc tgcagctcga
     2881 ggctcttccg tcaatcgagt tcaagggcga cacaaaattt attctaaatg cataataaat
     2941 actgataaca tcttatagtt tgtattatat tttgtattat cgttgacatg tataattttg
     3001 atatcaaaaa ctgattttcc ctttattatt ttcgagattt attttcttaa ttctctttaa
     3061 caaactagaa atattgtata tacaaaaaat cataaataat agatgaatag tttaattata
     3121 ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt
     3181 tataaggtta aataattctc atatatcaag caaagtgaca ggcgccctta aatattctga
     3241 caaatgctct ttccctaaac tccccccata aaaaaacccg ccgaagcggg tttttacgtt
     3301 atttgcggat taacgattac tcgttatcag aaccgcccag ggggcccgag cttaagactg
     3361 gccgtcgttt tacaacacag aaagagtttg tagaaacgca aaaaggccat ccgtcagggg
     3421 ccttctgctt agtttgatgc ctggcagttc cctactctcg ccttccgctt cctcgctcac
     3481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3961 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag
     4081 aagaacagta tttggt
//
