LOCUS       sequence-385210-        4657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector to introduce AID tag in N-terminal of the human 
            RPB1 gene. Auxin-inducible degron system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4657)
  AUTHORS   Valton AL, Venev SV, Mair B, Khokhar ES, Tong AHY, Usaj M, Chan K, 
            Pai AA, Moffat J, Dekker J
  TITLE     A cohesin traffic pattern genetically linked to gene regulation.
  JOURNAL   Nat Struct Mol Biol. 2022 Dec 8. doi: 10.1038/s41594-022-00890-9.
  PUBMED    36482254
REFERENCE   2  (bases 1 to 4657)
  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 Struct 
            Mol Biol. 2022 Dec 8. doi: 10.1038/s41594-022-00890-9."
COMMENT     Sequence Label: 221-AID-RPB1
FEATURES             Location/Qualifiers
     source          1..4657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     81..98
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(257..284)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(376..462)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     511..533
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     525..542
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     526..542
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1634..1765
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     primer_bind     complement(2887..2906)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2888..2906)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2911..2927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2911..2927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3029..3048)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3040..3849
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3107..3126)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3996..4584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4485..4504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
       61 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      121 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      181 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      241 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      301 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      361 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      421 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      481 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      541 gtcttaagct atctgctttt ctcggattgt cgcggtcccg tgatctctcc cggacagtgg
      601 ccaccctcct tccagagcag gcctcccagc tgaagtctct tcgagaggct caggccatgg
      661 atctaggatc atggctcggt acagcgcggc attacaggcc aggagatgcc cacccgtctc
      721 cgggggccgt ttgacaaaca tcggctactt ccggacaagc cactggtcct gccatcttgg
      781 tttgttcatg catattcagc aagagaactg catattcatg agcaaccgcc cttccccggg
      841 aggacccgcc ccgccccgcc gccattaccg attcgcagag cagaccattt tccagcttgg
      901 gggaggctac gaatgagagg cttttgctga agatgaaacc gttgtccaaa ctagaatgaa
      961 agcggcccaa ttttaccctc gagtcttcaa accttaagcc tcagtgcaaa agggagcagc
     1021 gaggagaggc ggtgggatta aagaaaaaaa ttcttctctc cctgtcactt caaaaggtcg
     1081 cccaaccagc gagcgggggg tttacccacg actctggctc gcaccccttg cgggcctgcg
     1141 caggaggccc cgtagcgcaa gggagggcgg gaaaggaagg ggcggggacg cgagggcgaa
     1201 tctataagaa gcgtcgttca gcgagttcgc tgctcagaag cgccgagagc gcggccggga
     1261 cggttggaga agaaggcggc tcccggaagg gggagagaca aactgccgta acctctgccg
     1321 ttcaggaacc cggttactta tttattcgtt accctttttc ttcttcctcc cccaaaaacc
     1381 ttttcctttt cccttctttt tttttccttt ttgggagctg aaaaatttcc ggtaagggaa
     1441 agaagggctc ctttcgctcc ttatttcccc gcctccttcc ctcccccacc ttcccctcct
     1501 ccggcttttt cctcccaact cggggaggtc cttcccggtg gccgccctga cgaggtctga
     1561 gcacctaggc ggaggcggcg caggcttttt gtagtgaggt ttgcgcctgc gcagcgcgcc
     1621 tgcctccgcc atgcctaaag atccagccaa acctccggcc aaggcacaag ttgtgggatg
     1681 gccaccggtg agatcatacc ggaagaacgt gatggtttcc tgccaaaaat caagcggtgg
     1741 cccggaggcg gcggcgttcg tgaagggggg agcaggaggc cacgggggtg gccccccctc
     1801 gggggacagc gcatgcccgc tgcgcaccat caagagagtc cagttcggag tcctgagtcc
     1861 ggatgaactg gtaagcggct ctgtcctccc cttcccccct cctcccctgg cgggcggggc
     1921 cggacggggg ctgcggaaac ttggcgcttt ctcgctgctt atgggtgacg ggccaggagc
     1981 atggctcagc agcgccaagg ccgtgtaggc gccagtctcg ggcctcccag agttatattt
     2041 tgcaaagagt tgagcaggcg tagcgctttg tccgagatgg ggggctggct ggagggtgag
     2101 aaggagggag agaaggatga ctgatttcac actccagagt taccgacgtt aaaggcgatc
     2161 cgacgaatcc tagggagtct tcacaagttt ggttaagaag ccaaacccgg gtagcttcct
     2221 cctcttctgt aatacaaatg ttaactgagc acctgctctg ttgaagacac aggagactcg
     2281 ggttacagag gtgaacacga cagaccagga ccctaccttc aagagcactg tctgtgggaa
     2341 agggcaagca gtaaacatga atacaatgaa tgaatgagat aattcagtta atggcatttg
     2401 tctcgaatag agtgaaaagg ggtaatccca tggagagtca ctgagctggg gctactttat
     2461 ttttttattt ttattttatt ttattttttg agacggagtc ttgctctgtc gcccaggctg
     2521 gagtgcaatg gcacgatctc ggctcactgc aatctccgcc tcccgggttc aagcgattct
     2581 cctgccttag cctcctgagt agctgggatt acaggcatgc gccaccatgc cccgctaatt
     2641 tttgtatttt tagtagagga ggggttttgc catgttggcc aggctggtct cgaacgcctg
     2701 ccctgaggtg attctccccc gtcttttttt tttttttttt tttttttttg agacggagtt
     2761 tcgctcttat tccccaggcc agagtgcaat ggctgaactc ctgacttcag gtgatccacc
     2821 cgcctcggcc tctcaaagtg ctgggattac aggcgtgagc tcattatttg ccatccagct
     2881 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2941 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     3001 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     3061 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     3121 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     3181 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     3241 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     3301 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3361 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     3421 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     3481 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     3541 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     3601 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     3661 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     3721 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     3781 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     3841 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     3901 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3961 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4021 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4081 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4141 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4201 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4261 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4321 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4381 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4441 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4501 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4561 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4621 ttttgctggc cttttgctca catgttcttt cctgcgt
//
