LOCUS       sequence-152675-        7140 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 7140)
  TITLE     Kate O'Connor-Giles Lab Donor Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7140)
  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            "Unpublished"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..7140
                     /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(7135..7140,1..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 gtacgtcatg ccccgttatt ctctattcgt
     4321 tttgtgactc tccctctctg tactattgct ctctcactct gtcgcacagt aaacggcact
     4381 ctattctcgt tgcttcgaga gagcgcgcct cgaatgttcg cgaaaagagc gccggagtat
     4441 aaatagaggc gcttcgtcga cggagcgtca attcaattca aacaagcaaa gtgaacacat
     4501 cgcgaagcaa gctgagcaaa caaacaagcg cagctgaaca agctaaacaa tctgcaataa
     4561 agtgcaagtt aaagtgaatc aattaaaagt aaccaacaac caagtaatta aactaaaaac
     4621 tgcaactact gaaatcaacc aagaagtcat tattgaagac aagaagagaa ctctgaatag
     4681 gtcgatagcg tcaattcggc acgaggtttc gtgacgaagc tccaagcggt ttacgccatc
     4741 aattaaacac aaagtgctgt gccaaaactc ctctcgcttc ttatttttgt ttgttttttg
     4801 agtgattggg tgggtgattg gttttgggtg ggtaagcagg ggaaagtgtg aaaaatcccg
     4861 gcaatgggcc aagaggatca ggagctatta attcgcggag gcagcaaaca cccatctgcc
     4921 gagcatctga acaatggtga cagcggagcg gcttcgcaga gctgcattaa ccagggcttc
     4981 gggcaggcca aaaactacgg cacgctcctg ccacccagtc cgccggagga ctccggttca
     5041 gggagcggcc aactagccga gaacctcacc tatgcctggc acaatatgga catctttggg
     5101 gcggtcaatc agccgggctc cggatggcgg cagctggtca accggacacg cggactattc
     5161 tgcaacgagc gacacatacc ggcgcccagg aaacatttgc tcaagaacgt ttgcggcgtg
     5221 gcctatccgg gcgaactttt ggccgtgatg ggcagttccg gtgccggaaa gacgaccctg
     5281 ctgaatgccc ttgcctttcg atcgccgcag ggcatccaag tatcgccatc cgggatgcga
     5341 ctgctcaatg gccaacctgt ggacgccaag gagatgcagg ccaggtgcgc ctatgtccag
     5401 caggatgacc tctttatcgg ctccctaacg gccagggaac acctgatttt ccaagccatg
     5461 gtgcggatgc cacgacatct gacctatcgg cagcgagtgg cccgcgtgga tcaggtgatc
     5521 caggagcttt cgctcagcaa atgtcagcac acgatcatcg gtgtgcccgg cagggtgaaa
     5581 ggtctgtccg gcggagaaag gaagcgtctg gcattcgcct ccgaggcact aaccgatccg
     5641 ccgcttctga tctgcgatga gcccacctcc ggactggact catttaccgc ccacagcgtc
     5701 gtccaggtgc tgaagaagct gtcgcagaag ggcaagaccg tcatcctgac cattcatcag
     5761 ccgtcttccg agctgtttga gctctttgac aagatccttc tgatggccga gggcagggta
     5821 gctttcttgg gcactcccag cgaagccgtc gacttctttt cctacgtggg tgcccagtgt
     5881 cctaccaact acaatccggc ggacttttac gtacaggtgt tggccgttgt gcccggacgg
     5941 gagatcgagt cccgtgatcg gatcgccaag atatgcgaca attttgccat tagcaaagta
     6001 gcccgggata tggagcagtt gttggccacc aaaaatctgg agaagccact ggagcagccg
     6061 gagaatgggt acacctacaa ggccacctgg ttcatgcagt tccgggcggt cctgtggcga
     6121 tcctggctgt cggtgctcaa ggaaccactc ctcgtaaaag tgcgacttat tcagacaacg
     6181 atggttgcca tcttgattgg cctcatcttt ttgggccaac aactcacgca agtgggtgtg
     6241 atgaatatca acggagccat cttcctcttc ctgaccaaca tgacctttca aaacgtcttt
     6301 gccacgataa atgtgttcac ctcagagctg ccagttttta tgagggaggc ccgaagtcga
     6361 ctttatcgct gtgacacata ctttctgggc aaaacgattg ccgaattgcc gctttttctc
     6421 acagtgccac tggtcttcac ggcgattgcc tatccgatga tcggactgcg ggccggagtg
     6481 ctgcacttct tcaactgcct ggcgctggtc actctggtgg ccaatgtgtc aacgtccttc
     6541 ggatatctaa tatcctgcgc cagctcctcg acctcgatgg cgctgtctgt gggtccgccg
     6601 gttatcatac cattcctgct ctttggcggc ttcttcttga actcgggctc ggtgccagta
     6661 tacctcaaat ggttgtcgta cctctcatgg ttccgttacg ccaacgaggg tctgctgatt
     6721 aaccaatggg cggacgtgga gccgggcgaa attagctgca catcgtcgaa caccacgtgc
     6781 cccagttcgg gcaaggtcat cctggagacg cttaacttct ccgccgccga tctgccgctg
     6841 gactacgtgg gtctggccat tctcatcgtg agcttccggg tgctcgcata tctggctcta
     6901 agacttcggg cccgacgcaa ggagtagccg acatatatcc gaaataactg cttgtttttt
     6961 ttttttacca ttattaccat cgtgtttact gtttattgcc ccctcaaaaa gctaatgtaa
     7021 ttatatttgt gccaataaaa acaagatatg acctatagaa tacaaaaaaa aaaaaaaaaa
     7081 aaaaaaaaaa aactcgacac cgggatccaa acctagcagc gtaatgctct gcttttacca
//
