LOCUS       sequence-152782-        7090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating dsDNA donors for HDR. Contains 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 7090)
  TITLE     Kate O'Connor-Giles Lab Donor Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7090)
  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            "Unpublished"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-w+
FEATURES             Location/Qualifiers
     source          1..7090
                     /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    1285..1318
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1544..1553
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1550..2227
                     /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(1680..1698)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1730..1750)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1948..1967
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2142..2165
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2348..2469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2385..2404)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2439..2458
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2476..2509
                     /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(3045..3062)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3045..3061)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3160..3177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3411..3430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7085..7090,1..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 gatcataact tcgtataatg tatgctatac gaagttatcg
     1321 tacgggatct aattcaatta gagactaatt caattagagc taattcaatt aggatccaag
     1381 cttatcgatt tcgaaccctc gaccgccgga gtataaatag aggcgcttcg tctacggagc
     1441 gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa
     1501 caagcgcagc tgaacaagct aaacaatcgg ctcgaagccg gtcgccacca tggcctcctc
     1561 cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     1621 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1681 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     1741 gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg actacaagaa
     1801 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     1861 ggtgaccgtg acccaggact cctccctcca ggacggctcc ttcatctaca aggtgaagtt
     1921 catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagacta tgggctggga
     1981 ggcgtccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccacaaggc
     2041 cctgaagctg aaggacggcg gccactacct ggtggagttc aagtccatct acatggccaa
     2101 gaagcccgtg cagctgcccg gctactacta cgtggactcc aagctggaca tcacctccca
     2161 caacgaggac tacaccatcg tggagcagta cgagcgcgcc gagggccgcc accacctgtt
     2221 cctgtagggg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2281 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2341 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2401 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2461 tgtatcttaa ccggtataac ttcgtataat gtatgctata cgaagttata gaagagcact
     2521 agtaaagatc tccatgcata aggcgcgcct aggccttctg cagctcgagg ctcttccgtc
     2581 aatcgagttc aagggcgaca caaaatttat tctaaatgca taataaatac tgataacatc
     2641 ttatagtttg tattatattt tgtattatcg ttgacatgta taattttgat atcaaaaact
     2701 gattttccct ttattatttt cgagatttat tttcttaatt ctctttaaca aactagaaat
     2761 attgtatata caaaaaatca taaataatag atgaatagtt taattatagg tgttcatcaa
     2821 tcgaaaaagc aacgtatctt atttaaagtg cgttgctttt ttctcattta taaggttaaa
     2881 taattctcat atatcaagca aagtgacagg cgcccttaaa tattctgaca aatgctcttt
     2941 ccctaaactc cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta
     3001 acgattactc gttatcagaa ccgcccaggg ggcccgagct taagactggc cgtcgtttta
     3061 caacacagaa agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag
     3121 tttgatgcct ggcagttccc tactctcgcc ttccgcttcc tcgctcactg actcgctgcg
     3181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtgggc taactacggc tacactagaa gaacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgacgcg cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc
     4021 aagtcgtacg actagcacct gactgtcgag cccagtggaa acccttgaaa tgcctttaag
     4081 tcgagcccag tggaaaccct tgaaatgcct ttaagtcgag cccagtggaa acccttgaaa
     4141 tgcctttaag tcgagcccag tggaaaccct tgaaatgcct ttaagtcgag cccagtggaa
     4201 acccttgaaa tgcctttaag tcgagtagca gtcgaggtag gtacgtcatg ccccgttatt
     4261 ctctattcgt tttgtgactc tccctctctg tactattgct ctctcactct gtcgcacagt
     4321 aaacggcact ctattctcgt tgcttcgaga gagcgcgcct cgaatgttcg cgaaaagagc
     4381 gccggagtat aaatagaggc gcttcgtcga cggagcgtca attcaattca aacaagcaaa
     4441 gtgaacacat cgcgaagcaa gctgagcaaa caaacaagcg cagctgaaca agctaaacaa
     4501 tctgcaataa agtgcaagtt aaagtgaatc aattaaaagt aaccaacaac caagtaatta
     4561 aactaaaaac tgcaactact gaaatcaacc aagaagtcat tattgaagac aagaagagaa
     4621 ctctgaatag gtcgatagcg tcaattcggc acgaggtttc gtgacgaagc tccaagcggt
     4681 ttacgccatc aattaaacac aaagtgctgt gccaaaactc ctctcgcttc ttatttttgt
     4741 ttgttttttg agtgattggg tgggtgattg gttttgggtg ggtaagcagg ggaaagtgtg
     4801 aaaaatcccg gcaatgggcc aagaggatca ggagctatta attcgcggag gcagcaaaca
     4861 cccatctgcc gagcatctga acaatggtga cagcggagcg gcttcgcaga gctgcattaa
     4921 ccagggcttc gggcaggcca aaaactacgg cacgctcctg ccacccagtc cgccggagga
     4981 ctccggttca gggagcggcc aactagccga gaacctcacc tatgcctggc acaatatgga
     5041 catctttggg gcggtcaatc agccgggctc cggatggcgg cagctggtca accggacacg
     5101 cggactattc tgcaacgagc gacacatacc ggcgcccagg aaacatttgc tcaagaacgt
     5161 ttgcggcgtg gcctatccgg gcgaactttt ggccgtgatg ggcagttccg gtgccggaaa
     5221 gacgaccctg ctgaatgccc ttgcctttcg atcgccgcag ggcatccaag tatcgccatc
     5281 cgggatgcga ctgctcaatg gccaacctgt ggacgccaag gagatgcagg ccaggtgcgc
     5341 ctatgtccag caggatgacc tctttatcgg ctccctaacg gccagggaac acctgatttt
     5401 ccaagccatg gtgcggatgc cacgacatct gacctatcgg cagcgagtgg cccgcgtgga
     5461 tcaggtgatc caggagcttt cgctcagcaa atgtcagcac acgatcatcg gtgtgcccgg
     5521 cagggtgaaa ggtctgtccg gcggagaaag gaagcgtctg gcattcgcct ccgaggcact
     5581 aaccgatccg ccgcttctga tctgcgatga gcccacctcc ggactggact catttaccgc
     5641 ccacagcgtc gtccaggtgc tgaagaagct gtcgcagaag ggcaagaccg tcatcctgac
     5701 cattcatcag ccgtcttccg agctgtttga gctctttgac aagatccttc tgatggccga
     5761 gggcagggta gctttcttgg gcactcccag cgaagccgtc gacttctttt cctacgtggg
     5821 tgcccagtgt cctaccaact acaatccggc ggacttttac gtacaggtgt tggccgttgt
     5881 gcccggacgg gagatcgagt cccgtgatcg gatcgccaag atatgcgaca attttgccat
     5941 tagcaaagta gcccgggata tggagcagtt gttggccacc aaaaatctgg agaagccact
     6001 ggagcagccg gagaatgggt acacctacaa ggccacctgg ttcatgcagt tccgggcggt
     6061 cctgtggcga tcctggctgt cggtgctcaa ggaaccactc ctcgtaaaag tgcgacttat
     6121 tcagacaacg atggttgcca tcttgattgg cctcatcttt ttgggccaac aactcacgca
     6181 agtgggtgtg atgaatatca acggagccat cttcctcttc ctgaccaaca tgacctttca
     6241 aaacgtcttt gccacgataa atgtgttcac ctcagagctg ccagttttta tgagggaggc
     6301 ccgaagtcga ctttatcgct gtgacacata ctttctgggc aaaacgattg ccgaattgcc
     6361 gctttttctc acagtgccac tggtcttcac ggcgattgcc tatccgatga tcggactgcg
     6421 ggccggagtg ctgcacttct tcaactgcct ggcgctggtc actctggtgg ccaatgtgtc
     6481 aacgtccttc ggatatctaa tatcctgcgc cagctcctcg acctcgatgg cgctgtctgt
     6541 gggtccgccg gttatcatac cattcctgct ctttggcggc ttcttcttga actcgggctc
     6601 ggtgccagta tacctcaaat ggttgtcgta cctctcatgg ttccgttacg ccaacgaggg
     6661 tctgctgatt aaccaatggg cggacgtgga gccgggcgaa attagctgca catcgtcgaa
     6721 caccacgtgc cccagttcgg gcaaggtcat cctggagacg cttaacttct ccgccgccga
     6781 tctgccgctg gactacgtgg gtctggccat tctcatcgtg agcttccggg tgctcgcata
     6841 tctggctcta agacttcggg cccgacgcaa ggagtagccg acatatatcc gaaataactg
     6901 cttgtttttt ttttttacca ttattaccat cgtgtttact gtttattgcc ccctcaaaaa
     6961 gctaatgtaa ttatatttgt gccaataaaa acaagatatg acctatagaa tacaaaaaaa
     7021 aaaaaaaaaa aaaaaaaaaa aactcgacac cgggatccaa acctagcagc gtaatgctct
     7081 gcttttacca
//
