LOCUS       sequence-474012-        6473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for Chd4-miniAID fusion, Roxed Bact-Bsd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6473)
  TITLE     Hendrich lab plasmids batch1
REFERENCE   2  (bases 1 to 6473)
  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: pChd4 GGSG_mAID-3xF_BactBsd
FEATURES             Location/Qualifiers
     source          1..6473
                     /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"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, 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"
     protein_bind    2281..2305
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3415..3618
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRDYK"
     CDS             3610..3675
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    3688..3719
                     /label=rox
                     /bound_moiety="Dre recombinase"
     CDS             4260..4658
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4781..4915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4818..4837)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4872..4891
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    4931..4962
                     /label=rox
                     /bound_moiety="Dre recombinase"
     primer_bind     complement(5832..5851)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5833..5851)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5858..5875)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5858..5874)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5867..5889)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6015..6470)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6152..6173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6364..6383
                     /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 ttcgatcggt accaaggcgc ataacgatac cacgatatca
     2281 acaagtttgt acaaaaaagc aggctggcgc cggaaccaat tcagtcgacg tttgaataag
     2341 ccggccaatg cagtcctaca caaaggtcgc cagtggctcc cctgcctcct gctccttcca
     2401 cctccccctc cactctcccc tccccaacat ccataaacag cttatttcca aactgtctgc
     2461 cgagcctgtg tggaacctta ggatgtgagc tgaggggatt ttctttgcag gcaatcgagg
     2521 acagcgattt tctttgaact cttaagtcta ttgagatttt gagcatagct ttatgggggg
     2581 ggggtgaaat gaaaaggtgg gtgtcataaa ggtagccggg gacattgggg atataaaaag
     2641 gtgtggatgc agagaggaga tggtaggaac tgagcctctc aagtggagga gcccaaggct
     2701 gagaataaaa actctgtggg catttgctgt gctcccaact tgggagcact tggaggcggg
     2761 agcacgatga gatgtgctca tgcttggctg cataggaagt ttgaagccaa ccttttatat
     2821 aaaaaaaaaa aaaaaaaaaa aaaaaaagaa agaaagaaaa agaaaatgga gacaatagtg
     2881 agaaggaata atgaactaaa gtttggtggc tcttaaaaac aagaatccgc cagctgtgga
     2941 tggccgatgc ctagagtcac agggactcaa gagacacagc aggacctgag ctcaagtctc
     3001 aggccatcct ggacaacatg tgagaccaga gccacttgca actagaatgg gaggaatagc
     3061 tgggtgggca ggccaggtaa agcgagtgtc aagagcgtat cctgtgtatg ccagcgtgac
     3121 aaccagccct gtgggccaca gctgtgccct tgactaccac agctcacggc ctctttcttt
     3181 ctttttccct ctgcagtcct gaaacagctg gaggaactgt tgagtgacat gaaagctgat
     3241 gtgacgcggc tcccagccac cattgcccgc atccccccag tcgctgtgag gttacagatg
     3301 tcagagcgga acattctcag ccgcctggca aacagggcgc cagaacctcc cccacagcag
     3361 gtagcccagc agcagaaacc cggctctggt ggcggcggaa gcgcctttga ttccaaggag
     3421 aagagtgctt gtcctaaaga tccagccaaa cctccggcca aggcacaagt tgtgggatgg
     3481 ccaccggtga gatcataccg gaagaacgtg atggtttcct gccaaaaatc aagcggtggc
     3541 ccggaggcgg cggcgttcgt gaaggtatca atggacggag caccgtactt gaggaaaatc
     3601 gatttgaggg actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag
     3661 gatgacgatg acaagtagaa acgatcgtaa ctttaaataa tgccaattat ttaaagttac
     3721 atatggcctt ctgcaggagc gtacagaacc cagggccctg gcacccgtgc agaccctggc
     3781 ccaccccacc tgggcgctca gtgcccaaga gatgtccaca cctaggatgt cccgcggtgg
     3841 gtggggggcc cgagagacgg gcaggccggg ggcaggcctg gccatgcggg gcctaaccgg
     3901 gcactgccca gcgtggggcg cgggggccac ggcgcgcgcc cccagccccc gggcccagca
     3961 ccccaaggcg gccaacgcca aaactctccc tcctcctctt cctcaatctc gctctcgctc
     4021 tttttttttt tcgcaaaagg aggggagagg gggtaaaaaa atgctgcact gtgcggcgaa
     4081 gccggtgagt gagcggcgcg gggccaatca gcgtgcgccg ttccgaaagt tgccttttat
     4141 ggctcgagcg gccgcggcgg cgccctataa aacccagcgg cgcgacgcgc caccaccgcc
     4201 gagaccgcgt ccgccccgcg agcacagagc ctcgcctttg ccgatccatc gatggatcca
     4261 tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca
     4321 acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc
     4381 gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgcgca gaactcgtgg
     4441 tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa
     4501 atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggttctt ctcgatctgc
     4561 atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc
     4621 gtgaattgct gccctctggt tatgtgtggg agggctaagc tctagataag taatgatcat
     4681 aatcagccat atcacatctg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4741 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4801 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4861 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccattt
     4921 actgatcact taactttaaa taatgccaat tatttaaagt taggcgcgag agtgatatcc
     4981 acctccacca ctcagcagtg accttcctca ctttctcttg tcccggctgc ccctgggggc
     5041 cagagagacc cttgccttcc ttctgcccat cttctgagtt gtaaaggaac agccctgcgc
     5101 agtggggcag ggagggaggg agaagcaatg gtgcccttcc tgctggagag acaagcagca
     5161 gtagcgcagc gcccgcccgc catgtgcagg agctggcggg ctggctgcct tctggaccct
     5221 ggcttctccc caccgtgcgc ctgttacaaa ctgttgtgga ttctgccagg ctcgaagaaa
     5281 atgatctgaa tttcttcctc tttttggttt tattttgttg gtttactttg tgttttctcc
     5341 tttttggggg gtattcagag tgggccgggc ccctgggcga gacacagcta cctctgttgg
     5401 catcttttta ataccaggaa cccagcggct ccagccactg agcagctaaa taaagttgga
     5461 aaaaaaaaaa aaagaaaaag aaccaaaagc ataaaaaacc acagcaaatt tcttgatgag
     5521 aactgaaaat aaaagtttcc ttgtatttta gtgatctggt tctgtgtggg tgagtgtggg
     5581 ttccaggtac cccaggaact ggcttatcct ggaagactgc ctgtgctcct gacagggtga
     5641 cattctggcg aacaagaggc ttctggacca ttgtttatag gctctgatct ttagcaaact
     5701 gttcgcagag cactcctgca ggaagtagcc acttgcctgg tacagttgtg aagacattgg
     5761 gcaagtgttc cgatatctag acccagcttc tagttctaga gcggccgcca ccgcggtgga
     5821 gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     5881 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5941 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6001 tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     6061 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6121 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6181 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6241 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6301 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6361 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6421 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cag
//
