LOCUS       sequence-205829-        7090 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  HPRT1 c.312C>A (Munich) donor vector with bilateral microhomology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7090)
  AUTHORS   Kim SI, Matsumoto T, Kagawa H, Nakamura M, Hirohata R, Ueno A, 
            Ohishi M, Sakuma T, Soga T, Yamamoto T, Woltjen K
  TITLE     Microhomology-assisted scarless genome editing in human iPSCs.
  JOURNAL   Nat Commun. 2018 Mar 5;9(1):939. doi: 10.1038/s41467-018-03044-y.
  PUBMED    29507284
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: "Nat 
            Commun."; date: "2018-03-5"; volume: "9(1):939. doi"; pages: " 
            10.1038/s41467-018-03044-y"
COMMENT     Sequence Label: p3-HPRT1-S104Rf-PdTK-mCh
FEATURES             Location/Qualifiers
     source          1..7090
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     8..27
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    94..118
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             124..834
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(269..287)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     537..556
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     731..754
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(836..860)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(956..975)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1021..1076
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1075..1094)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     1849..1866
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1850..1866
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1887..1910)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     primer_bind     complement(1990..2007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2161..2749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2241..2260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2920..3711,3712..3780))
                     /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     3543..3562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3781..3885)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3953..3971
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4009..4031)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4131..4150
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             5149..5202
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5212..5808
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5212..5231)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5708..5728
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5815..6810
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(6842..6859)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6848..7072
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgcggcc
       61 gcggtaccgg cgcgccggat cctgcaggtt atcacaagtt tgtacaaaaa agcaggctcc
      121 accatggtga gcaagggcga ggaggacaac atggccatca tcaaggagtt catgcgcttc
      181 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      241 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
      301 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
      361 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      421 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      481 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
      541 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
      601 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
      661 gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
      721 gtcaacatca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
      781 gagcgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaccca
      841 gctttcttgt acaaagtggt gataactcta gagaattcac tcctcaggtg caggctgcct
      901 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
      961 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     1021 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     1081 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     1141 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     1201 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     1261 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     1321 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     1381 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggatcc ctcgagttaa
     1441 ttaaatctag aagtcgacgg gcacgggcag cttgccggtg gtaagagata ttgtgtgagt
     1501 atatttaata tatgattctt tttagtggca acagtaggtt ttcttatatt ttctttgaat
     1561 ctctgcaaac catacttgct ttcatttcac ttggttacag tgagattttt ctaacatatt
     1621 cactagtact ttacatcaaa gccaatactg tttttttaaa actagtcacc ttggaggata
     1681 tatacttatt ttacaggtgt gtgtggtttt ttaaataaac tccttttagg aattgctgtt
     1741 gggacttggg atactttttt cactatacat actggtgaca gataccctct cttgagctac
     1801 atcggtttgt ggggagtcaa aagtcctttg gagctaggtt tgacaaattg taaaacgacg
     1861 gccagtgcgg ccgcgaattc ggtaccctaa ctaacaggaa gagtttgtag gggccccccc
     1921 tcgactgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1981 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2041 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2101 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2161 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2221 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2281 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2341 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2401 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2461 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2521 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2581 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2641 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2701 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2761 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2821 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2881 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2941 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3001 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3061 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3121 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3181 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3241 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3301 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3361 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3421 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3481 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3541 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3601 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3661 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3721 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3781 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3841 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3901 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3961 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4021 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4081 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4141 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     4201 cgatcgcgaa aataccgcat caggcgtgca gtgcagcaga atgatcacag ttcactgcag
     4261 tctcaacctc ctgggttcaa gcgatccttc cacctcagcc tcctgagtag ctgggactat
     4321 aggtgtgcgc caccactccc agctaatatt tttattttgt agaaacaggg ttcgccatgt
     4381 tacccagcct cccaaagtgc tgggattaca ggcatgagcc actggcccag tttctacagt
     4441 ctctcttaat attgtatatt atccaagaaa tttcatttaa tcagaacctg ccagtctgat
     4501 aggtgaaaat ggtatcttgt ttttatttgc atttaaaaaa aattatgata gtggtatgct
     4561 tggttttttt gaaggtatca aattttttac cttatgaaac atgagggcaa aggatgtgtt
     4621 acgtggaaga tttaaaaaaa atttttaatg catttttttg agacaaggtc ttgctctatt
     4681 gtccaggctg gagtgcagtg gcacaatcac agttcactcc agcctcaaca tcctgcacta
     4741 aagtgatttt cccacctcac ctctcaagta gctgggacta caggtacatg ctaccatgcc
     4801 tggctaattt tttttttttt gcaggcatgg ggtctcacta tattgcccag gttggtgtgg
     4861 aagtttaatg actaagaggt gtttgttata aagtttaatg tatgaaactt tctattaaat
     4921 tcctgatttt atttctgtag gactgaacgt cttgctcgag atgtgatgaa ggagatggga
     4981 ggccatcaca ttgtagccct ctgtgtgctc aaggggggct ataaattctt tgctgacctg
     5041 ctggattaca tcaaagcact gaatagaaat agtgatagat ccattcctat gactgtagat
     5101 tttatcagac ttaagagata ttgtaccacc ggcaagctgc ccgtgcccga gggcagagga
     5161 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctaggctcga gatgaccgag
     5221 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     5281 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     5341 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5401 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5461 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5521 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5581 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5641 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5701 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5761 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgg atccatgccc
     5821 acgctactgc gggtttatat agacggtccc cacgggatgg ggaaaaccac caccacgcaa
     5881 ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg tacccgagcc gatgacttac
     5941 tggcgggtgc tgggggcttc cgagacaatc gcgaacatct acaccacaca acaccgcctc
     6001 gaccagggtg agatatcggc cggggacgcg gcggtggtaa tgacaagcgc ccagataaca
     6061 atgggcatgc cttatgccgt gaccgacgcc gttctggctc ctcatatcgg gggggaggct
     6121 gggagctcac atgccccgcc cccggccctc accctcatct tcgaccgcca tcccatcgcc
     6181 gccctcctgt gctacccggc cgcgcggtac cttatgggca gcatgacccc ccaggccgtg
     6241 ctggcgttcg tggccctcat cccgccgacc ttgcccggca ccaacatcgt gcttggggcc
     6301 cttccggagg acagacacat cgaccgcctg gccaaacgcc agcgccccgg cgagcggctg
     6361 gacctggcta tgctggctgc gattcgccgc gtttacgggc tacttgccaa tacggtgcgg
     6421 tatctgcagt gcggcgggtc gtggcgggag gactggggac agctttcggg gacggccgtg
     6481 ccgccccagg gtgccgagcc ccagagcaac gcgggcccac gaccccatat cggggacacg
     6541 ttatttaccc tgtttcgggc ccccgagttg ctggccccca acggcgacct gtataacgtg
     6601 tttgcctggg ccttggacgt cttggccaaa cgcctccgtt ccatgcacgt ctttatcctg
     6661 gattacgacc aatcgcccgc cggctgccgg gacgccctgc tgcaacttac ctccgggatg
     6721 gtccagaccc acgtcaccac ccccggctcc ataccgacga tatgcgacct ggcgcgcacg
     6781 tttgcccggg agatggggga ggctaactga tctagagggc ccgtttaaac ccgctgatca
     6841 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6901 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6961 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     7021 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggactagcgg
     7081 tggcggcctc
//
