LOCUS       sequence-342536-        7078 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DDA1 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7078)
  AUTHORS   Faust TB, Yoon H, Nowak RP, Donovan KA, Li Z, Cai Q, Eleuteri NA, 
            Zhang T, Gray NS, Fischer ES
  TITLE     Structural complementarity facilitates E7820-mediated degradation of
            RBM39 by DCAF15.
  JOURNAL   Nat Chem Biol. 2020 Jan;16(1):7-14. doi: 10.1038/s41589-019-0378-3. 
            Epub 2019 Nov 4.
  PUBMED    31686031
REFERENCE   2  (bases 1 to 7078)
  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"; doi: 
            "10.1038/s41589-019-0378-3"; journalName: "Nat Chem Biol"; date: 
            "2020-01"; volume: "16"; issue: "1"; pages: "7-14"
COMMENT     Sequence Label: pTF023_His_hsDDA1
FEATURES             Location/Qualifiers
     source          1..7078
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1344..1799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1431..1450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1641..1662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1816..1835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1935..1957
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1995..2013)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2081..2185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2186..2254,2255..3046)
                     /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(2404..2423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3217..3805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3706..3725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3959..3976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_recomb     4368..5195
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4368..4492
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4530..5135)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    complement(5199..5232)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5422..5470
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5495..5514
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5548..6225)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Strongin et al., 2007)"
                     /label=DsRed-Monomer
                     /note="mammalian codon-optimized"
                     /translation="MDNTEDVIKEFMQFKVRMEGSVNGHYFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYMKLSFPEGFTWERSMNFEDGGV
                     VEVQQDSSLQDGTFIYKVKFKGVNFPADGPVMQKKTAGWEPSTEKLYPQDGVLKGEISH
                     ALKLKDGGHYTCDFKTVYKAKKPVQLPGNHYVDSKLDITNHNEDYTVVEQYEHAEARHS
                     GSQ"
     primer_bind     complement(5808..5827)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(6234..6343)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6398..6489
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     6414..6431
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             6513..6530
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6543..6563
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_recomb     join(6918..7078,1..1197)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(join(6934..7078,1..1197))
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
ORIGIN
        1 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc
       61 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct
      121 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg
      181 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat
      241 tgactcgacg taaacacgtt aaataaagct tggacatatt taacatcggg cgtgttagct
      301 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
      361 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
      421 tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc gggtttcaat
      481 ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc aggcggtggt
      541 aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga taaatctacc
      601 atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg tggcggtgat
      661 gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac ctcaactatt
      721 gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg atttttttcg
      781 tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg ttgaggttcc
      841 gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg tggtggaggc
      901 gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat aatttgttct
      961 ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg ctgcacaacg
     1021 gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt ataattggaa
     1081 tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac cgtgccgata
     1141 tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa gctcggatcg
     1201 atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt ggcgagacac
     1261 ttcgctgtcg tcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     1321 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1381 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1441 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1501 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     1561 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1621 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1681 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     1741 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     1801 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     1861 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     1921 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     1981 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     2041 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2101 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2161 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2221 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2281 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2341 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2401 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2461 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2521 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2581 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2641 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2701 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2761 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2821 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2881 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2941 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3001 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3061 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3121 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3181 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3241 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3301 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3361 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3421 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3481 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3541 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3601 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3661 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3721 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3781 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3841 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3901 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3961 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4021 ttggccgatt cattaatgca gaacggctcc gcccactatt aatgaaatta aaaattccaa
     4081 ttttaaaaaa cgcagcaaga gaaacatttg tatgaaagaa tgcgtagaag gaaagaaaaa
     4141 tgtcgtcgac atgctgaaca acaagattaa tatgcctccg tgtataaaaa aaatattgaa
     4201 cgatttgaaa gaaaacaatg taccgcgcgg cggtatgtac aggaagaggt ttatactaaa
     4261 ctgttacatt gcaaacgtgg tttcgtgtgc caagtgtgaa aaccgatgtt taatcaaggc
     4321 tctgacgcat ttctacaacc acgactccaa gtgtgtgggt gaagtcatgc atcttttaat
     4381 caaatcccaa gatgtgtata aaccaccaaa ctgccaaaaa atgaaaactg tcgacaagct
     4441 ctgtccgttt gctggcaact gcaagggtct caatcctatt tgtaattatt gaataataaa
     4501 acaattataa atgtcaaatt tgttttttat taacgataca aaccaaacgc aacaagaaca
     4561 tttgtagtat tatctataat tgaaaacgcg tagttataat cgctgaggta atatttaaaa
     4621 tcattttcaa atgattcaca gttaatttgc gacaatataa ttttattttc acataaacta
     4681 gacgccttgt cgtcttcttc ttcgtattcc ttctcttttt catttttctc ctcataaaaa
     4741 ttaacatagt tattatcgta tccatatatg tatctatcgt atagagtaaa ttttttgttg
     4801 tcataaatat atatgtcttt tttaatgggg tgtatagtac cgctgcgcat agtttttctg
     4861 taatttacaa cagtgctatt ttctggtagt tcttcggagt gtgttgcttt aattattaaa
     4921 tttatataat caatgaattt gggatcgtcg gttttgtaca atatgttgcc ggcatagtac
     4981 gcagcttctt ctagttcaat tacaccattt tttagcagca ccggattaac ataactttcc
     5041 aaaatgttgt acgaaccgtt aaacaaaaac agttcacctc ccttttctat actattgtct
     5101 gcgagcagtt gtttgttgtt aaaaataaca gccattgtaa tgagacgcac aaactaatat
     5161 cacaaactgg aaatgtctat caatatatag ttgctgatat aacttcgtat agcatacatt
     5221 atacgaagtt atctggttga aacgtacccg tagtggctat ggcagggctt gccgccccga
     5281 cgttggctgc gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag
     5341 aaacgcgggc gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct
     5401 gggaccgaac cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt
     5461 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     5521 ctcccccatc tcccgctatt actcgagtca ctgggagccg gagtggcggg cctcggcgtg
     5581 ctcgtactgc tccaccacgg tgtagtcctc gttgtggttg gtgatgtcca gcttggagtc
     5641 cacgtagtgg ttgccgggca gctgcacggg cttcttggcc ttgtacacgg tcttgaagtc
     5701 gcaggtgtag tggccgccgt ccttcagctt cagggcgtgg gagatctcgc ccttcagcac
     5761 gccgtcctgg gggtacagct tctcggtgga gggctcccag ccggcagtct tcttctgcat
     5821 tacggggccg tcggcgggga agttcacgcc cttgaacttc accttgtaga tgaaggtgcc
     5881 gtcctgcagg gaggagtcct gctgcacctc caccacgccg ccgtcctcga agttcatgga
     5941 gcgctcccag gtgaagccct cggggaagga cagcttcatg tagtcgggga tgtcggcggg
     6001 gtgcttcacg taggccttgg agccgtactg gaactggggg gacaggatgt cccaggcgaa
     6061 gggcaggggg ccgcccttgg tcacctgcag cttggcggtc tgggtgccct cgtagggctt
     6121 gccctcgccc tcgccctcga tctcgaagta gtggccgttc acggagccct ccatgcgcac
     6181 cttgaactgc atgaactcct tgatgacgtc ctcggtgttg tccatggtag agtgattgta
     6241 aataaaatgt aatttacagt atagtatttt aattaatata caaatgattt gataataatt
     6301 cttatttaac tataatatat tgtgttgggt tgaattaaag gtccgtatac tagtatcgat
     6361 tcgcgaccta ctccggaata ttaatagagt tgctgatatc atggagataa ttaaaatgat
     6421 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
     6481 cctataaata cggatctacc atgggcagca gccatcatca tcatcatcac tccgcggtcg
     6541 acgagaacct ctacttccag ggaggcggcc gcatggcgga tttcctgaag ggattgccgg
     6601 tttataacaa aagtaacttt agcagatttc acgcagattc agtctgcaag gcgtccaaca
     6661 ggcgcccgag cgtttacctt ccgactcgtg agtatccttc cgaacagatc atcgtcacgg
     6721 aaaaaacgaa cattcttctt agatacctgc atcagcagtg ggacaaaaaa aatgccgcaa
     6781 aaaaaagaga tcaggagcaa gttgagctgg aaggtgagtc cagtgcgccg ccgcgcaaag
     6841 tggcgaggac tgatagccct gatatgcacg aagacacgta gggtaccccc ggacggccgc
     6901 ttaattaatt gatccgggtt attagtacat ttattaagcg ctagattctg tgcgttgttg
     6961 atttacagac aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg
     7021 gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatatt
//
