LOCUS       sequence-426857-        4147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HDR template to tag endogenous human RANGAP1 C-terminus with 
            miniIAA7.3xFlag.P2A.BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4147)
  AUTHORS   Li S, Wang Y, van der Stoel M, Zhou X, Madhusudan S, Kanerva K, 
            Nguyen VD, Eskici N, Olkkonen VM, Zhou Y, Raivio T, Ikonen E
  TITLE     HiHo-AID2: boosting homozygous knock-in efficiency enables robust 
            generation of human auxin-inducible degron cells.
  JOURNAL   Genome Biol. 2024 Feb 26;25(1):58. doi: 10.1186/s13059-024-03187-w.
  PUBMED    38409044
REFERENCE   2  (bases 1 to 4147)
  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: "Genome 
            Biol."; date: "2024-02-26"; volume: "25(1):58. doi"; pages: " 
            10.1186/s13059-024-03187-w"
COMMENT     Sequence Label: pGL3-RANGAP.C-linker.miniIAA7.3xFlag.P2A.BSD
FEATURES             Location/Qualifiers
     source          1..4147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(73..90)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(244..832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(324..343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1003..1794,1795..1863))
                     /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     1626..1645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1864..1968)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2608..2673
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2683..2739
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
ORIGIN
        1 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
       61 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca cgccagtcct
     1981 atgtggccac tggcagccag taaggattcc ctggcctatg tctgctggaa gggcccttgg
     2041 agagcctggc tgagggggcc ttcattctgg ccagctctgg gctgtgtggc gttgggtgcc
     2101 tggctctctg agcctctcgc acctcccatc cccacccacg gtgtgcagaa ggcagagggc
     2161 taatgtctct cttgctgagt cgttgtgcaa attaacaaaa atgagtctgg caccaaaatg
     2221 gcccccacac ttggaaaaca ggccaagagg catccaccga cctgctgacc cctctttccc
     2281 cgcaggccca acagcgccct ggaatcctgc tccttcgccc gccacagtct gctgcagacg
     2341 ctgtacaaag tcggatctgg cggaggaggg agcgggggag gcggatctgg cggaggcgga
     2401 tccggcttct ctgagaccgt ggacctgatg ctgaacctgc agtccaataa ggagggctct
     2461 gtggatctga agaacgtgag cgccgtgcct aaggagaaga ccacactgaa ggacccatcc
     2521 aagccccctg ccaaggcaca ggtggtggga tggccacccg tgcggaacta cagaaagaat
     2581 atgatgaccc agcagaagac aagctccgac tacaaagacc atgacggtga ttataaagat
     2641 catgacatcg attacaagga tgacgatgac aagggctccg gcgccaccaa cttttctctg
     2701 ctgaagcagg ccggcgatgt ggaggagaat cctggcccag gagccaagcc actgagccag
     2761 gaggagagca cactgattga aagagcaact gccactatta acagcattcc cattagcgag
     2821 gactattctg tggcaagcgc cgccctgagc tccgacggaa ggatcttcac cggcgtgaac
     2881 gtgtaccact ttacaggcgg cccctgcgcc gagctggtgg tgctgggaac cgcagcagca
     2941 gcagcagcag gcaatctgac atgtatcgtg gccatcggca acgagaatag gggcatcctg
     3001 tccccatgcg gccggtgtag acaggtgctg ctggatctgc accctggcat caaggccatc
     3061 gtgaaggact ctgatggaca gccaaccgcc gtcgggatta gagaactgct gccatcagga
     3121 tacgtctggg agggctagtg tacaagtaga ctcaaagcct ctcccatccc ttggcctgga
     3181 ccagtgagct ggggagggac tcggatgaac tgaggcgcag cctacgccat tgccttggac
     3241 aggactctgg ccacaggcag ggcgggtctg tgtcccatgt gtcctgtcag tcccctgagt
     3301 atgtgtgtgg gtgtggcgca tgtgcaggtc tgtgcctcct gtcgggattt gggttttaac
     3361 gtcttctgct ggcccagccc tgctctgttg tggggagttg gcccccaggg gaaagggctg
     3421 tgagctgctc cgccattaaa ctcacctcca cctgagggcg ctctgctgat ctccgcctgg
     3481 gccctgatgg ccgtccccac ccacctgcct tccggcccgg ctccctggcg gagccagaac
     3541 ccaaggagtt gcccgcgtgc tgtccttccc ctctgtgttg tgattgggtt gtttcctgcc
     3601 ctgcctgggg ctgcttctcg tcaccaagcc ctggtcctgc ggcagctgtc acccctacca
     3661 tccataccac tgtgctgacc gctcagcctg aagagcagag aatgccatgg gtgggactgt
     3721 gggggtcgga tcgtggggtt gttggcagag ggcaaccctg ggccccacac cgtgtggaca
     3781 ggcagacacc agattgtcca ggagcaggag ctgctgggac tgcgctggcc ccggacctag
     3841 tgggccttct cctggctgct gagatgtcgt ctgtgactgg cctggctgga gggggagtgt
     3901 tgacaaccca aagctgttct ccagtctggg gagggagagg cagggtcccc aatgtccgag
     3961 ctgcatctgg acgctgctct taaaggacct cctggggcag gggagcggta gggtctggac
     4021 tgggcagatg ctgtatgacc tccctgagca cccgtgactg ccccatgctt tcccctttgt
     4081 gctctgtgtg tgtctgggct gtgcccgggg gcttcacaaa taaagtcgtg tggcagcttc
     4141 agagact
//
