LOCUS       sequence-227021-        6452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA-tagged mVenus fused to mAID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6452)
  AUTHORS   Daniel K, Icha J, Horenburg C, Muller D, Norden C, Mansfeld J
  TITLE     Conditional control of fluorescent protein degradation by an 
            auxin-dependent nanobody.
  JOURNAL   Nat Commun. 2018 Aug 17;9(1):3297. doi: 10.1038/s41467-018-05855-5.
  PUBMED    30120238
REFERENCE   2  (bases 1 to 6452)
  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: "2018-08-17"; volume: "9(1):3297. doi"; pages: " 
            10.1038/s41467-018-05855-5"
COMMENT     Sequence Label: pIREShygro HA-mVenus-mAID
FEATURES             Location/Qualifiers
     source          1..6452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        508..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     756..775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        756..774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             832..858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             862..888
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             892..918
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(919..921,922..1632)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(961..982)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1222..1241)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1569..1590
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1651..1782
                     /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"
     intron          1911..2140
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2123..2142
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2198..2774
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2368..2385)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2595..2614
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2810..3835
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2810..3835
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    3965..4086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4002..4021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4188..4204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4188..4204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4201..4223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4212..4228
                     /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(4236..4242,4243..4260,4261..4266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4281..4302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4419..4436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4590..5178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4670..4689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5349..6140,6141..6209))
                     /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     5972..5991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6210..6314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6389..6408)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
       61 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      121 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      481 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      541 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      601 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      721 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      781 caagcttggt accgagctcg gatcgatatc tgcggcctag ctagcatggc ctacccatac
      841 gatgttcctg actatgcggg ctatccctat gacgtcccgg actatgcagg ctatccatat
      901 gacgttccag attacgctgt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      961 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1021 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     1081 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     1141 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1201 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1261 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1321 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     1381 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1441 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1501 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     1561 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1621 gagctgtaca agggatccgg aggtgcctgt cccaaagatc cagccaagcc cccagccaag
     1681 gcccaagtgg tggggtggcc cccagtaagg tcctaccgca agaatgtcat ggtcagttgt
     1741 caaaagtcct ctggtggacc tgaagcagcc gctttcgtga aagtgtcaat ggatggagct
     1801 ccatacctga gaaagattga cctgagaatg tataagtgat aggcggccgc atagataact
     1861 gatccagtgt gctggaatta attcgctgtc tgcgagggcc agctgttggg gtgagtactc
     1921 cctctcaaaa gcgggcatga cttctgcgct aagattgtca gtttccaaaa acgaggagga
     1981 tttgatattc acctggcccg cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga
     2041 aaagacaatc tttttgttgt caagcttgag gtgtggcagg cttgagatct ggccatacac
     2101 ttgagtgaca atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     2161 ctgcaggtcg agcatgcatc tagggcggcc aattccgccc ctctcccccc cccccccccc
     2221 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     2281 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     2341 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     2401 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     2461 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     2521 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     2581 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     2641 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     2701 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     2761 aaacacgatg ataagcttgc cacaacccgg gataattcct gcagccaata tgaaaaagcc
     2821 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     2881 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     2941 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     3001 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     3061 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     3121 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     3181 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     3241 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     3301 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     3361 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     3421 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     3481 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     3541 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     3601 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     3661 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     3721 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     3781 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataggggga
     3841 tcaattctct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3901 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3961 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4081 atcttaacgc gtcgagtgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4141 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4201 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4261 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4321 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5341 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5401 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5461 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5521 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5581 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5641 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5701 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5761 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5821 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5881 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5941 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6001 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6061 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6121 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6181 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6241 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6301 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6361 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6421 agtatctgct ccctgcttgt gtgttggagg tc
//
