LOCUS       sequence-234561-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  AUTHORS   Yesbolatova A, Natsume T, Hayashi KI, Kanemaki MT
  TITLE     Generation of conditional auxin-inducible degron (AID) cells and 
            tight control of degron-fused proteins using the degradation 
            inhibitor auxinole.
  JOURNAL   Methods. 2019 Apr 23. pii: S1046-2023(18)30331-1. doi: 
            10.1016/j.ymeth.2019.04.010.
  PUBMED    31026591
REFERENCE   2  (bases 1 to 5457)
  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: "Methods. 
            2019 Apr 23. pii: S1046-2023(18)30331-1. doi: 
            10.1016/j.ymeth.2019.04.010."
COMMENT     Sequence Label: pMK389 (mCherry-BSD)
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..106
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     98..115
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     99..115
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     125..144
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        125..143
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             182..892
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(327..345)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    1010..1131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1047..1066)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1101..1120
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1162..1195
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1256..1755
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1291..1314)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1688..1707
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1779..1788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1785..2183
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    2686..2719
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2768..2788)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2768..2786)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2807..2823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2807..2823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2820..2842)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2831..2847
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2855..2861,2862..2879,2880..2885))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2900..2921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3038..3055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3209..3797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3289..3308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3968..4759,4760..4828))
                     /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     4591..4610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4829..4933)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4960..5415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5047..5066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5257..5278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
       61 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
      121 cgcgtaatac gactcactat agggcgaatt ggagctcgga tccggtgcag gcgccgctag
      181 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
      241 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
      301 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
      361 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
      421 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca attgggagcg
      481 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
      541 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
      601 aatgcagtgt cgtaccatgg gctgggaggc ctccactgag cggatgtacc ccgaggacgg
      661 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
      721 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
      781 cgacatcaag ttggacatcc tttcccacaa cgaggactac accatcgtgg aacagtacga
      841 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aatctagata
      901 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
      961 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1021 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1081 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtcga
     1141 tcatattcaa taacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt
     1201 ctgaagagga gtttacgtcc agccaagctt aggatctcga cctcgaaatt ctaccgggta
     1261 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     1321 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     1381 ccgactccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     1441 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     1501 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     1561 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     1621 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     1681 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     1741 tctccgggcc tttcgacctg catccatcta gatctcgagc caccatggcc aagcctttgt
     1801 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     1861 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     1921 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     1981 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     2041 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     2101 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     2161 ctggttatgt gtgggagggc taatcgatga tatcagatcc ccgggatgca gaaattgatg
     2221 atctattaaa caataaagat gtccactaaa atggaagttt ttcctgtcat actttgttaa
     2281 gaagggtgag aacagagtac ctacattttg aatggaagga ttggagctac gggggtgggg
     2341 gtggggtggg attagataaa tgcctgctct ttactgaagg ctctttacta ttgctttatg
     2401 ataatgtttc atagttggat atcataattt aaacaagcaa aaccaaatta agggccagct
     2461 cattcctccc actcatgatc tatagatcta tagatctctc gtgggatcat tgtttttctc
     2521 ttgattccca ctttgtggtt ctaagtactg tggtttccaa atgtgtcagt ttcatagcct
     2581 gaagaacgag atcagcagcc tctgttccac atacacttca ttctcagtat tgttttgcca
     2641 agttctaatt ccatcagaag ctggtcgaga tccggaaccc ttaatataac ttcgtataat
     2701 gtatgctata cgaagttatt aggtccctcg aagaggttca ctaggatccg gtacccagct
     2761 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     2821 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2881 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2941 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3961 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4021 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4081 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4141 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4201 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4261 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4321 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4381 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4441 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4501 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4561 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4621 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4681 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4741 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4801 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4861 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4921 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4981 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5041 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5101 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5161 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5221 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5281 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5341 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5401 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatc
//
