LOCUS       sequence-474013-        8404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting construct to make C-term fusion with GGSG-AID-3xFLAG, 
            roxed Pgk-Puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8404)
  TITLE     Hendrich lab plasmids batch1
REFERENCE   2  (bases 1 to 8404)
  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"
COMMENT     Sequence Label: pMbd3-GGSG-AID-3xF_PP
FEATURES             Location/Qualifiers
     source          1..8404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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     complement(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2217..2233
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2218..2234
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3360..4043
                     /codon_start=1
                     /gene="Arabidopsis thaliana AXR3"
                     /product="auxin-responsive IAA17 protein"
                     /label=auxin-inducible degron
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="MGSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNN
                     EPANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSG
                     GPEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMN
                     ERKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAME
                     KCKSRA"
     CDS             4044..4109
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    4157..4181
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4219..4250
                     /label=rox
                     /bound_moiety="Dre recombinase"
     promoter        4300..4799
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4335..4358)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4732..4751
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      4813..4822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4819..5418
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5318..5338
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5425..6420
                     /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="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    6448..6672
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    6720..6751
                     /label=rox
                     /bound_moiety="Dre recombinase"
     primer_bind     complement(7763..7782)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7764..7782)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7789..7806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7789..7805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7798..7820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(7946..8401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8083..8104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8295..8314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagctcga aattaaccct cactaaaggg aacaaaagct ggtaccgggc cccccctcga
     2221 ggtcgacggt atcgataagc ttgatatcgg gcctgggatg gggtcataac ttccaggtag
     2281 agccctaggc ccaagggttc ctctgaccct gcccactctc atattccaca ggagtgggcc
     2341 ctggctgtac agatgagacg ctgctgtcag ccattgcgag tgctctacac accagcaccc
     2401 tgcccattac aggccagctc tctgcagccg tggagaagaa ccctggtgtg tggctgaaca
     2461 ctgcacagcc actgtgcaaa gccttcatgg tgacagatga cgacatcagg taactggcag
     2521 ctctgccccc accccaatat ctccacagtt agcacaagaa tgtagggcat gccagcaact
     2581 acccggcccc tcaggagtgg tctgtgagtg aaggaaattt agatgcccga caggcacgct
     2641 tagaaatgct gggttgggga aatagcagtg agaaagcatg gcttccacag gcctggggga
     2701 gctgcacaca gagcagaaga tgggtggggc caaactgagt gagtgacagc cctcaggagt
     2761 gtgggggggg ggggatggtg gaccaggaag aaggcagctt aaactacctc tgagctgcag
     2821 tctggaccct cggggctagc ctgaatttcc cttgtcagac ttggtgatca ggccagatgg
     2881 ggacagggca ggacccagaa ttaggttcta gatgcacagt cgaggctctc cttgctatgt
     2941 gagcatggcc cgtagctggt cttactctgc cttcccatag cttgcatgta aaaaagtcag
     3001 aatgtaagga gaccatggtc acccaaggtg acagagaagc tgtgtggagt ttctgaaagg
     3061 ggctgtggtg atgcgcttgt ggactggtcc ctgtgtgccc tgtcagagtg tgttctccta
     3121 ggaagcagga ggagctggta cagcaggtac ggaagcgcct ggaggaggca ctgatggccg
     3181 acatgctagc tcatgtggag gagcttgccc gagacgggga ggcaccactg gacaaggcct
     3241 gtgcagagga ggaagaggag gaggaagagg aggaggaaga gccggagcca gagcgagtgg
     3301 gctctggtgg cggtggctct ggcggaggtg ggtccggtgg cggcggaagc gccccaggaa
     3361 tgggcagtgt cgagctgaat ctgagggaga ctgagctgtg tcttggtctt cccggtggag
     3421 atacagtggc tccggtaacc ggaaacaaga gagggttctc agagacggtt gatctgaagc
     3481 taaatctgaa taatgagcct gcaaacaagg aaggatctac gactcatgac gtcgtgactt
     3541 ttgattccaa ggagaagagt gcttgtccta aagatccagc caaacctccg gccaaggcac
     3601 aagttgtggg atggccaccg gtgagatcat accggaagaa cgtgatggtt tcctgccaaa
     3661 aatcaagcgg tggcccggag gcggcggcgt tcgtgaaggt atcaatggac ggagcaccgt
     3721 acttgaggaa aatcgatttg aggatgtata aaagctacga tgagctttct aatgctttgt
     3781 ccaacatgtt cagctctttt accatgggca aacatggagg agaagaagga atgatagact
     3841 tcatgaatga gaggaaattg atggatttgg tgaatagctg ggactatgtt ccctcttatg
     3901 aagacaaaga cggtgattgg atgctcgtcg gcgacgttcc ttggccaatg ttcgtcgata
     3961 catgcaagcg tttacgtctc atgaaaggat cggatgccat tggtctcgct ccgagggcga
     4021 tggagaagtg caagagcaga gctgactaca aagaccatga cggtgattat aaagatcatg
     4081 acatcgacta caaggatgac gatgacaagt agaagcttcg atcggtacca aggcgcataa
     4141 cgataccacg atatcaacaa gtttgtacaa aaaagcaggc tggcgccgga accaattcag
     4201 tcgacgttta aacgatcgta actttaaata atgccaatta tttaaagtta catatgagaa
     4261 agtataggaa cttcagcttg atatcgaatt aattctaccg ggtaggggag gcgcttttcc
     4321 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     4381 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     4441 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     4501 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     4561 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     4621 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     4681 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     4741 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     4801 cctgcagcca acgccaccat ggggaccgag tacaagccca cggtgcgcct cgccacccgc
     4861 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4921 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4981 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5041 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5101 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5161 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5221 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5281 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5341 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5401 acccgcaagc ccggtgccgg atccatgccc acgctactgc gggtttatat agacggtcct
     5461 cacgggatgg ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat
     5521 atcgtctacg tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc
     5581 gcgaacatct acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg
     5641 gcggtggtaa tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc
     5701 gttctggctc ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc
     5761 accctcatct tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac
     5821 cttatgggca gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc
     5881 ttgcccggca caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg
     5941 gccaaacgcc agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc
     6001 gtttacgggc tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag
     6061 gattggggac agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac
     6121 gcgggcccac gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg
     6181 ctggccccca acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa
     6241 cgcctccgtc ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg
     6301 gacgccctgc tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc
     6361 ataccgacga tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga
     6421 gctctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6481 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6541 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     6601 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     6661 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgacatac tagagtactt
     6721 aactttaaat aatgccaatt atttaaagtt aggcgcgccg atatctagac ccagcttcta
     6781 gttctagagc ttagcacagg tgctgcaggc cagggtggga tgtcagccca tcccctagcc
     6841 gggctgctaa cagcttccct tcctgcctat tgcaggtgcc ctgcccaagt ctgggctgca
     6901 gactgccttc agccttgcct ggaccaggta ggggccagac ctgtaggagg cagccgtcca
     6961 cctcctttcc aaagcctcct gcttccaggt ctcagtgcag ggagcccctg tggaccttga
     7021 actcacttgt ccctgcgctg cctggcagga agccccacac tgaaagcaga tgagcagtga
     7081 cccaactgag aggccacctg gacacagtca cctccctgcc tccttatcat aggacaaggc
     7141 cttgcttggc accgaggagc tgggagccgt gttgggtgct ggaggaagtt tctggaaaca
     7201 cacctggcta tgcccacctt atgtccctaa ggctattaca ggccagggtc tggactgctc
     7261 cggcccacag ggctgcccag cctccccaca ctgagggtca gcagcccacc aggaagtcac
     7321 tttccttcaa taaactgatg gtaggaactt gtgaactctg acttgtcact gtgtgccaca
     7381 cctctgaggg tgtctgctgc ccgggcccat ggaaacctgc tttgggcctt agtgtccctg
     7441 tgtggcaggg acacaggctc caactttgag agagcagttg gcagcctctc tgtgcttggt
     7501 gcctagatct tggacacctg aactgtgccc ctgcagtcgg agcctccaag aaactgacgg
     7561 gtgagctggg gtgtgtagcc cagtgagcga gtcagtctgt ggatacaccc cagtgactga
     7621 gctcggccac cctaaatcag atcagggtct tgttccccag cctagacagc actcagggct
     7681 ggagtggtgc cttttgtcct catcccaaaa ggaccaccca gctaccctgt cacatgggcc
     7741 accgcggtgg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     7801 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     7861 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     7921 gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg
     7981 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     8041 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     8101 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     8161 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     8221 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     8281 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     8341 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     8401 tcag
//
