LOCUS       sequence-172486-        7144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating dsDNA donors for HDR. Contains an attP docking
            site, the visible marker 3xP3-DsRed and GMR-white for identifying 
            backbone incorporation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7144)
  TITLE     Kate O'Connor-Giles Lab Donor Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7144)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pHD-DsRed-attP-w+
FEATURES             Location/Qualifiers
     source          1..7144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     618..637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(856..947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     989..1008
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1014..1033
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1335..1368
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1594..1603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1600..2277
                     /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(1730..1748)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1780..1800)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1998..2017
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2192..2215
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2398..2519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2435..2454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2489..2508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2526..2559
                     /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(3095..3112)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3095..3111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3210..3227)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3381..3969)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3461..3480)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7139..7144,1..786,787..855))
                     /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 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
       61 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagcgc
      121 tgcgatgata ccgcgagaac cacgctcacc ggctccggat ttatcagcaa taaaccagcc
      181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      301 tgccatcgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      601 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
      721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
      781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
      841 atgttgaata ctcatattct tcctttttca atattattga agcatttatc agggttattg
      901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggtcagtgt
      961 tacaaccaat taaccaattc tgaacattat cgcgagccca tttatacctg aatatggctc
     1021 ataacacccc ttgtttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1081 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gactccccat gcgagagtag
     1141 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgcccg
     1201 ggctaattat ggggtgtcgc ccttcgctga agcaggtgga attcttgcat gctagcggcc
     1261 gcggacatat gcacacctgc gatcgtagtg ccccaactgg ggtaaccttt gagttctctc
     1321 agttgggggc gtagataact tcgtataatg tatgctatac gaagttatcg tacgggatct
     1381 aattcaatta gagactaatt caattagagc taattcaatt aggatccaag cttatcgatt
     1441 tcgaaccctc gaccgccgga gtataaatag aggcgcttcg tctacggagc gacaattcaa
     1501 ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc
     1561 tgaacaagct aaacaatcgg ctcgaagccg gtcgccacca tggcctcctc cgaggacgtc
     1621 atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc
     1681 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1741 gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac
     1801 ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc
     1861 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1921 acccaggact cctccctcca ggacggctcc ttcatctaca aggtgaagtt catcggcgtg
     1981 aacttcccct ccgacggccc cgtaatgcag aagaagacta tgggctggga ggcgtccacc
     2041 gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccacaaggc cctgaagctg
     2101 aaggacggcg gccactacct ggtggagttc aagtccatct acatggccaa gaagcccgtg
     2161 cagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca caacgaggac
     2221 tacaccatcg tggagcagta cgagcgcgcc gagggccgcc accacctgtt cctgtagggg
     2281 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2341 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2401 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2461 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2521 ccggtataac ttcgtataat gtatgctata cgaagttata gaagagcact agtaaagatc
     2581 tccatgcata aggcgcgcct aggccttctg cagctcgagg ctcttccgtc aatcgagttc
     2641 aagggcgaca caaaatttat tctaaatgca taataaatac tgataacatc ttatagtttg
     2701 tattatattt tgtattatcg ttgacatgta taattttgat atcaaaaact gattttccct
     2761 ttattatttt cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata
     2821 caaaaaatca taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc
     2881 aacgtatctt atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat
     2941 atatcaagca aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc
     3001 cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc
     3061 gttatcagaa ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa
     3121 agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct
     3181 ggcagttccc tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3241 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3301 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3361 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3421 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3481 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3541 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3601 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3661 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3721 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3781 gagttcttga agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc
     3841 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3901 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
     4021 cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc aagtcgtacg
     4081 actagcacct gactgtcgag cccagtggaa acccttgaaa tgcctttaag tcgagcccag
     4141 tggaaaccct tgaaatgcct ttaagtcgag cccagtggaa acccttgaaa tgcctttaag
     4201 tcgagcccag tggaaaccct tgaaatgcct ttaagtcgag cccagtggaa acccttgaaa
     4261 tgcctttaag tcgagtagca gtcgaggtag gtacgccatg ccccgttatt ctctattcgt
     4321 tttgtgactc tccctctctg tactattgct ctctcactct gtcgcacagt aaacggcacg
     4381 ctattctcgt tgcttcgaga gagcgcgcct cgaatgttcg cgaaaagagc gccggagtat
     4441 aaatagaggc gcttcgtcga cggagcgtca attcaattca aacaagcaaa gtgaacacat
     4501 cgcgaagcgt aagctgagca aacaaacaag cgcagctgaa caagctaaac aatctgcaat
     4561 aaagtgcaag ttaaagtgaa tcaattaaaa gtaaccaaca accaagtaat taaactaaaa
     4621 actgcaacta ctgaaatcaa ccaagaagtc attattgaag acaagaagag aactctgaat
     4681 aggtcgatag cgtcaattcg gcacgagcgg tttcgtgacg aagctccaag cggtttacgc
     4741 catcaattaa acacaaagtg ctgtgccaaa actcctctcg cttcttattt ttgtttgttt
     4801 tttgagtgat tgggtgggtg attggttttg ggtgggtaag caggggaaag tgtgaaaaat
     4861 cccggcaatg ggccaagagg atcaggagct attaattcgc ggaggcagca aacacccatc
     4921 tgccgagcat ctgaacaatg gtgacagcgg agcggcttcg cagagctgca ttaaccaggg
     4981 cttcgggcag gccaaaaact acggcacgct cctgccaccc agtccgccgg aggactccgg
     5041 ttcagggagc ggccaactag ccgagaacct cacctatgcc tggcacaata tggacatctt
     5101 tggggcggtc aatcagccgg gctccggatg gcggcagctg gtcaaccgga cacgcggact
     5161 attctgcaac gagcgacaca taccggcgcc caggaaacat ttgctcaaga acgtttgcgg
     5221 cgtggcctat ccgggcgaac ttttggccgt gatgggcagt tccggtgccg gaaagacgac
     5281 cctgctgaat gcccttgcct ttcgatcgcc gcagggcatc caagtatcgc catccgggat
     5341 gcgactgctc aatggccaac ctgtggacgc caaggagatg caggccaggt gcgcctatgt
     5401 ccagcaggat gacctcttta tcggctccct aacggccagg gaacacctga ttttccaggc
     5461 catggtgcgg atgccacgac atctgaccta tcggcagcga gtggcccgcg tggatcaggt
     5521 gatccaggag ctttcgctca gcaaatgtca gcacacgatc atcggtgtgc ccggcagggt
     5581 gaaaggtctg tccggcggag aaaggaagcg tctggcattc gcctccgagg cactaaccga
     5641 tccgccgctt ctgatctgcg atgagcccac ctccggactg gactcattta ccgcccacag
     5701 cgtcgtccag gtgctgaaga agctgtcgca gaagggcaag accgtcatcc tgaccattca
     5761 tcagccgtct tccgagctgt ttgagctctt tgacaagatc cttctgatgg ccgagggcag
     5821 ggtagctttc ttgggcactc ccagcgaagc cgtcgacttc ttttcctacg tgggtgccca
     5881 gtgtcctacc aactacaatc cggcggactt ttacgtacag gtgttggccg ttgtgcccgg
     5941 acgggagatc gagtcccgtg atcggatcgc caagatatgc gacaattttg ccattagcaa
     6001 agtagcccgg gatatggagc agttgttggc caccaaaaat ttggagaagc cactggagca
     6061 gccggagaat gggtacacct acaaggccac ctggttcatg cagttccggg cggtcctgtg
     6121 gcgatcctgg ctgtcggtgc tcaaggaacc actcctcgta aaagtgcgac ttattcagac
     6181 aacgatggtt gccatcttga ttggcctcat ctttttgggc caacaactca cgcaagtggg
     6241 cgtgatgaat atcaacggag ccatcttcct cttcctgacc aacatgacct ttcaaaacgt
     6301 ctttgccacg ataaatgtgt tcacctcaga gctgccagtt tttatgaggg aggcccgaag
     6361 tcgactttat cgctgtgaca catactttct gggcaaaacg attgccgaat tgccgctttt
     6421 tctcacagtg ccactggtct tcacggcgat tgcctatccg atgatcggac tgcgggccgg
     6481 agtgctgcac ttcttcaact gcctggcgct ggtcactctg gtggccaatg tgtcaacgtc
     6541 cttcggatat ctaatatcct gcgccagctc ctctacctcg atggcgctgt ctgtgggtcc
     6601 gccggttatc ataccattcc tgctctttgg cggcttcttc ttgaactcgg gctcggtgcc
     6661 agtatacctc aaatggttgt cgtacctctc atggttccgt tacgccaacg agggtctgct
     6721 gattaaccaa tgggcggacg tggagccggg cgaaattagc tgcacatcgt cgaacaccac
     6781 gtgccccagt tcgggcaagg tcatcctgga gacgcttaac ttctccgccg ccgatctgcc
     6841 gctggactac gtgggtctgg ccattctcat cgtgagcttc cgggtgctcg catatctggc
     6901 tctaagactt cgggcccgac gcaaggagta gccgacatat atccgaaata actgtttgtt
     6961 tttttttttt accattatta ccatcgtgtt tactgtttat tgccccctca aaaagcttat
     7021 gtaattatat ttgtgccaat aaaaacaaga tatgacctat agaatacaaa aaaaaaaaaa
     7081 aaaaaaaaaa aaaaaactcg acaccgggat ccaaacctag cagcgtaatg ctctgctttt
     7141 acca
//
