LOCUS       sequence-76999-e        4098 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 4098)
  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 4098)
  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..4098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    1339..1372
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1598..1607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1604..2281
                     /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(1734..1752)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1784..1804)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2002..2021
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2196..2219
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2402..2523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2439..2458)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2493..2512
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2530..2563
                     /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(3099..3116)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3099..3115)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3214..3231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3385..3973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3465..3484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4097..4098,1..790,791..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 ctgcgatcgt agtgccccaa ctggggtaac ctttgagttc
     1321 tctcagttgg gggcgtagat aacttcgtat aatgtatgct atacgaagtt atcgtacggg
     1381 atctaattca attagagact aattcaatta gagctaattc aattaggatc caagcttatc
     1441 gatttcgaac cctcgaccgc cggagtataa atagaggcgc ttcgtctacg gagcgacaat
     1501 tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa taaacaagcg
     1561 cagctgaaca agctaaacaa tcggctcgaa gccggtcgcc accatggcct cctccgagga
     1621 cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     1681 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1741 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca
     1801 gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc
     1861 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1921 cgtgacccag gactcctccc tccaggacgg ctccttcatc tacaaggtga agttcatcgg
     1981 cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct gggaggcgtc
     2041 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca aggccctgaa
     2101 gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg ccaagaagcc
     2161 cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct cccacaacga
     2221 ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc tgttcctgta
     2281 ggggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2341 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2401 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2521 ttaaccggta taacttcgta taatgtatgc tatacgaagt tatagaagag cactagtaaa
     2581 gatctccatg cataaggcgc gcctaggcct tctgcagctc gaggctcttc cgtcaatcga
     2641 gttcaagggc gacacaaaat ttattctaaa tgcataataa atactgataa catcttatag
     2701 tttgtattat attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt
     2761 ccctttatta ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta
     2821 tatacaaaaa atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa
     2881 aagcaacgta tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc
     2941 tcatatatca agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa
     3001 actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt
     3061 actcgttatc agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac
     3121 agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat
     3181 gcctggcagt tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt
     3241 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3781 tacagagttc ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat
     3841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4021 cgcgcgcgta actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca
     4081 gcgtaatgct ctgctttt
//
