LOCUS       sequence-152801-        7611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating dsDNA donors for HDR. Contains the visible 
            marker 3xP3-DsRed flanked by piggyBac termini and GMR-white for 
            identifying backbone incorporation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7611)
  TITLE     Kate O'Connor-Giles Lab Donor Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7611)
  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            "Unpublished"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: pScarlessHD-DsRed-w+
FEATURES             Location/Qualifiers
     source          1..7611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      106..694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     595..614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     848..865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     963..980
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     964..980
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1526..1588
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      1983..1992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1989..2666
                     /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(2119..2137)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2169..2189)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2387..2406
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2581..2604
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2787..2908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2824..2843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2878..2897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3183..3217
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3513..3532)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3538..3557)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3599..3690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3691..3759,3760..4551)
                     /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     complement(3909..3928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc
       61 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
      121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      241 accaaatact gttcttctag tgtagccgta gttagcccac cacttcaaga actctgtagc
      301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      721 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      781 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      841 cgagcgcagc gagtcagtga gcgaggaagc ggaaggcgag agtagggaac tgccaggcat
      901 caaactaagc agaaggcccc tgacggatgg cctttttgcg tttctacaaa ctctttctgt
      961 gttgtaaaac gacggccagt cttaagctcg ggccccctgg gcggttctga taacgagtaa
     1021 tcgttaatcc gcaaataacg taaaaacccg cttcggcggg tttttttatg gggggagttt
     1081 agggaaagag catttgtcag aatatttaag ggcgcctgtc actttgcttg atatatgaga
     1141 attatttaac cttataaatg agaaaaaagc aacgcacttt aaataagata cgttgctttt
     1201 tcgattgatg aacacctata attaaactat tcatctatta tttatgattt tttgtatata
     1261 caatatttct agtttgttaa agagaattaa gaaaataaat ctcgaaaata ataaagggaa
     1321 aatcagtttt tgatatcaaa attatacatg tcaacgataa tacaaaatat aatacaaact
     1381 ataagatgtt atcagtattt attatgcatt tagaatacgt actcagcgtc tgggttcccc
     1441 atcggtgatg tcgtataaga gacgtatagg agacctatag tgtcttcggg gccgaattcg
     1501 caggtggcag cacctgcgat cttaacccta gaaagataat catattgtga cgtacgttaa
     1561 agataatcat gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
     1621 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
     1681 aacaaacaat ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct
     1741 tctataaagt aacaaaactt ttaggatcta attcaattag agactaattc aattagagct
     1801 aattcaatta ggatccaagc ttatcgattt cgaaccctcg accgccggag tataaataga
     1861 ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca cgtcgctaag
     1921 cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatcggc tcgaagccgg
     1981 tcgccaccat ggcctcctcc gaggacgtca tcaaggagtt catgcgcttc aaggtgcgca
     2041 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2101 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct
     2161 gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag caccccgccg
     2221 acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2281 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctccag gacggctcct
     2341 tcatctacaa ggtgaagttc atcggcgtga acttcccctc cgacggcccc gtaatgcaga
     2401 agaagactat gggctgggag gcgtccaccg agcgcctgta cccccgcgac ggcgtgctga
     2461 agggcgagat ccacaaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca
     2521 agtccatcta catggccaag aagcccgtgc agctgcccgg ctactactac gtggactcca
     2581 agctggacat cacctcccac aacgaggact acaccatcgt ggagcagtac gagcgcgccg
     2641 agggccgcca ccacctgttc ctgtagcggc cgcgactcta gatcataatc agccatacca
     2701 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2761 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2821 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2881 gtttgtccaa actcatcaat gtatcttaga tatctataac aagaaaatat atatataata
     2941 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     3001 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     3061 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     3121 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     3181 tgcatgcgtc aattttacgc agactatctt tctagggtta aggaagagcc gtcgctcttc
     3241 cccgggaaaa gaagaccata tacgtctcct ataggtctcg tatactgctt cctatacgac
     3301 atcaccgatg gggaacacga agcgatccag cccccctaat tagcccgggc gaaaggccca
     3361 gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcatggg
     3421 gagtccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc atggggtcag
     3481 gtgggaccac cgcgctactg ccgccaggca aacaaggggt gttatgagcc atattcaggt
     3541 ataaatgggc tcgcgataat gttcagaatt ggttaattgg ttgtaacact gacccctatt
     3601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3661 atgcttcaat aatattgaaa aaggaagaat atgagtattc aacatttccg tgtcgccctt
     3721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4201 ataccaaacg acgagcgtga caccacgatg cctgtagcga tggcaacaac gttgcgcaaa
     4261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4381 gataaatccg gagccggtga gcgtggttct cgcggtatca tcgcagcgct ggggccagat
     4441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta aaagcagagc
     4561 attacgctgc taggtttgga tcccggtgtc gagttttttt tttttttttt tttttttttt
     4621 ttgtattcta taggtcatat cttgttttta ttggcacaaa tataattaca ttagcttttt
     4681 gagggggcaa taaacagtaa acacgatggt aataatggta aaaaaaaaaa acaagcagtt
     4741 atttcggata tatgtcggct actccttgcg tcgggcccga agtcttagag ccagatatgc
     4801 gagcacccgg aagctcacga tgagaatggc cagacccacg tagtccagcg gcagatcggc
     4861 ggcggagaag ttaagcgtct ccaggatgac cttgcccgaa ctggggcacg tggtgttcga
     4921 cgatgtgcag ctaatttcgc ccggctccac gtccgcccat tggttaatca gcagaccctc
     4981 gttggcgtaa cggaaccatg agaggtacga caaccatttg aggtatactg gcaccgagcc
     5041 cgagttcaag aagaagccgc caaagagcag gaatggtatg ataaccggcg gacccacaga
     5101 cagcgccatc gaggtcgagg agctggcgca ggatattaga tatccgaagg acgttgacac
     5161 attggccacc agagtgacca gcgccaggca gttgaagaag tgcagcactc cggcccgcag
     5221 tccgatcatc ggataggcaa tcgccgtgaa gaccagtggc actgtgagaa aaagcggcaa
     5281 ttcggcaatc gttttgccca gaaagtatgt gtcacagcga taaagtcgac ttcgggcctc
     5341 cctcataaaa actggcagct ctgaggtgaa cacatttatc gtggcaaaga cgttttgaaa
     5401 ggtcatgttg gtcaggaaga ggaagatggc tccgttgata ttcatcacac ccacttgcgt
     5461 gagttgttgg cccaaaaaga tgaggccaat caagatggca accatcgttg tctgaataag
     5521 tcgcactttt acgaggagtg gttccttgag caccgacagc caggatcgcc acaggaccgc
     5581 ccggaactgc atgaaccagg tggccttgta ggtgtaccca ttctccggct gctccagtgg
     5641 cttctccaga tttttggtgg ccaacaactg ctccatatcc cgggctactt tgctaatggc
     5701 aaaattgtcg catatcttgg cgatccgatc acgggactcg atctcccgtc cgggcacaac
     5761 ggccaacacc tgtacgtaaa agtccgccgg attgtagttg gtaggacact gggcacccac
     5821 gtaggaaaag aagtcgacgg cttcgctggg agtgcccaag aaagctaccc tgccctcggc
     5881 catcagaagg atcttgtcaa agagctcaaa cagctcggaa gacggctgat gaatggtcag
     5941 gatgacggtc ttgcccttct gcgacagctt cttcagcacc tggacgacgc tgtgggcggt
     6001 aaatgagtcc agtccggagg tgggctcatc gcagatcaga agcggcggat cggttagtgc
     6061 ctcggaggcg aatgccagac gcttcctttc tccgccggac agacctttca ccctgccggg
     6121 cacaccgatg atcgtgtgct gacatttgct gagcgaaagc tcctggatca cctgatccac
     6181 gcgggccact cgctgccgat aggtcagatg tcgtggcatc cgcaccatgg cttggaaaat
     6241 caggtgttcc ctggccgtta gggagccgat aaagaggtca tcctgctgga cataggcgca
     6301 cctggcctgc atctccttgg cgtccacagg ttggccattg agcagtcgca tcccggatgg
     6361 cgatacttgg atgccctgcg gcgatcgaaa ggcaagggca ttcagcaggg tcgtctttcc
     6421 ggcaccggaa ctgcccatca cggccaaaag ttcgcccgga taggccacgc cgcaaacgtt
     6481 cttgagcaaa tgtttcctgg gcgccggtat gtgtcgctcg ttgcagaata gtccgcgtgt
     6541 ccggttgacc agctgccgcc atccggagcc cggctgattg accgccccaa agatgtccat
     6601 attgtgccag gcataggtga ggttctcggc tagttggccg ctccctgaac cggagtcctc
     6661 cggcggactg ggtggcagga gcgtgccgta gtttttggcc tgcccgaagc cctggttaat
     6721 gcagctctgc gaagccgctc cgctgtcacc attgttcaga tgctcggcag atgggtgttt
     6781 gctgcctccg cgaattaata gctcctgatc ctcttggccc attgccggga tttttcacac
     6841 tttcccctgc ttacccaccc aaaaccaatc acccacccaa tcactcaaaa aacaaacaaa
     6901 aataagaagc gagaggagtt ttggcacagc actttgtgtt taattgatgg cgtaaaccgc
     6961 ttggagcttc gtcacgaaac ctcgtgccga attgacgcta tcgacctatt cagagttctc
     7021 ttcttgtctt caataatgac ttcttggttg atttcagtag ttgcagtttt tagtttaatt
     7081 acttggttgt tggttacttt taattgattc actttaactt gcactttatt gcagattgtt
     7141 tagcttgttc agctgcgctt gtttgtttgc tcagcttgct tcgcgatgtg ttcactttgc
     7201 ttgtttgaat tgaattgacg ctccgtcgac gaagcgcctc tatttatact ccggcgctct
     7261 tttcgcgaac attcgaggcg cgctctctcg aagcaacgag aatagagtgc cgtttactgt
     7321 gcgacagagt gagagagcaa tagtacagag agggagagtc acaaaacgaa tagagaataa
     7381 cggggcatga cgtacctacc tcgactgcta ctcgacttaa aggcatttca agggtttcca
     7441 ctgggctcga cttaaaggca tttcaagggt ttccactggg ctcgacttaa aggcatttca
     7501 agggtttcca ctgggctcga cttaaaggca tttcaagggt ttccactggg ctcgacttaa
     7561 aggcatttca agggtttcca ctgggctcga cagtcaggtg ctagtcgtac g
//
