LOCUS       sequence-443894-        4567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for generating dsDNA donors for homology-directed repair. 
            Contains the visible marker 3xP3-DsRed flanked by piggyBac termini..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4567)
  TITLE     pHD-pBac-DsRed
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4567)
  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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..4567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..174
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     206..224
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      351..360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   755..817
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1363..1380)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1363..1379)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1478..1495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1649..2237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1729..1748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2359..3150,3151..3219))
                     /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     2982..3001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3220..3311)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3353..3372
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3378..3397
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     repeat_region   3693..3727
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    4002..4123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4013..4032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4067..4086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(4244..4567,1..354))
                     /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(4306..4329)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4504..4523)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gaaggagccg tcctggaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa
       61 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcttgt agtcggggat
      121 gtcggcgggg tgcttcacgt acaccttgga gccgtactgg aactgggggg acaggatgtc
      181 ccaggcgaag ggcagggggc cgcccttggt caccttcagc ttggcggtct gggtgccctc
      241 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc
      301 catgcgcacc ttgaagcgca tgaactcctt gatgacgtcc tcggaggagg ccatggtggc
      361 gaccggcttc gagccgattg tttagcttgt tcagctgcgc ttgtttattt gcttagcttt
      421 cgcttagcga cgtgttcact ttgcttgttt gaattgaatt gtcgctccgt agacgaagcg
      481 cctctattta tactccggcg gtcgagggtt cgaaatcgat aagcttggat cctaattgaa
      541 ttagctctaa ttgaattagt ctctaattga attagatcct aaaagttttg ttactttata
      601 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
      661 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
      721 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
      781 ctttaacgta cgtcacaata tgattatctt tctagggtta agatcgcagg tgctgccacc
      841 tgcgaattcg gccccgaaga cactataggt ctcctatacg tctcttatac gacatcaccg
      901 atggggaacc cagacgctga gtacgtattc taaatgcata ataaatactg ataacatctt
      961 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
     1021 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
     1081 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     1141 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     1201 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     1261 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     1321 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     1381 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     1441 tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     1501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2041 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     2101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2281 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     2341 gtcagcgtaa tgctctgctt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2401 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2461 gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct caccggctcc
     2521 ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2581 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2641 agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc
     2701 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2761 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2821 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2881 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2941 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3001 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3061 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3121 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3181 aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt ttcaatatta
     3241 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3301 aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag
     3361 cccatttata cctgaatatg gctcataaca ccccttgttt gcctggcggc agtagcgcgg
     3421 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     3481 tggggactcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     3541 tcgaaagact gggcctttcg cccgggctaa taaggggggc tggatcgctt cgtgttcccc
     3601 atcggtgatg tcgtatagga agcagtatac gagacctata ggagacgtat atggtcttct
     3661 tttcccgggg aagagcgacg gctcttcctt aaccctagaa agatagtctg cgtaaaattg
     3721 acgcatgcat tcttgaaata ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc
     3781 atttaggaca tctcagtcgc cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa
     3841 gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa atgacgcatg attatctttt
     3901 acgtgacttt taagatttaa ctcatacgat aattatattg ttatttcatg ttctacttac
     3961 gtgataactt attatatata tattttcttg ttatagatat ctaagataca ttgatgagtt
     4021 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4081 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4141 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4201 ctacaaatgt ggtatggctg attatgatct agagtcgcgg ccgctacagg aacaggtggt
     4261 ggcggccctc ggcgcgctcg tactgctcca cgatggtgta gtcctcgttg tgggaggtga
     4321 tgtccagctt ggagtccacg tagtagtagc cgggcagctg cacgggcttc ttggccatgt
     4381 agatggactt gaactccacc aggtagtggc cgccgtcctt cagcttcagg gccttgtgga
     4441 tctcgccctt cagcacgccg tcgcgggggt acaggcgctc ggtggacgcc tcccagccca
     4501 tagtcttctt ctgcattacg gggccgtcgg aggggaagtt cacgccgatg aacttcacct
     4561 tgtagat
//
