LOCUS       sequence-449175-        9231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of  D3 and ASK1 complex in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9231)
  AUTHORS   Shabek N, Ticchiarelli F, Mao H, Hinds TR, Leyser O, Zheng N
  TITLE     Structural plasticity of D3-D14 ubiquitin ligase in strigolactone 
            signalling.
  JOURNAL   Nature. 2018 Nov;563(7733):652-656. doi: 10.1038/s41586-018-0743-5. 
            Epub 2018 Nov 21.
  PUBMED    30464344
REFERENCE   2  (bases 1 to 9231)
  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/s41586-018-0743-5"; journalName: "Nature"; date: "2018-11";
            volume: "563"; issue: "7733"; pages: "652-656"
COMMENT     Sequence Label: pFB-D3-ASK1
FEATURES             Location/Qualifiers
     source          1..9231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      184..638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(271..290)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     480..501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        664..768
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(769..837,838..1629)
                     /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(987..1006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1800..2388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2289..2308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2542..2559
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2693..2917
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2984..3517)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3687..3692,3693..3709,3710..3715))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        4063..4154
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             4221..4238
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4641..4658
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5040..5057
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5439..5456
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5463..5483
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6912..6932
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             7050..7070
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    7880..8014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        8092..8183
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             8216..8236
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     polyA_signal    8895..9029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  9066..9231
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca
       61 aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt
      121 cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat
      181 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag
      481 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      541 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      601 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
      661 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      721 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      781 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      841 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      901 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      961 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1021 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1081 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1141 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1201 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1261 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1321 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1381 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1441 gctggctggt ttattgctga taaatctgga gccggtgagc gtggctctcg cggtatcatt
     1501 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1561 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1621 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1681 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1741 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1801 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1861 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1921 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1981 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2041 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2101 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2161 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2221 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2281 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2341 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2401 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2461 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2521 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2581 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca gccgcgtaac
     2641 ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg
     2701 gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact
     2761 agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact
     2821 tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa
     2881 gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga
     2941 acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg
     3001 ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg
     3061 cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct
     3121 catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga
     3181 ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc agaacgtaag
     3241 ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac
     3301 tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt
     3361 ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc
     3421 cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac
     3481 tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     3541 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca
     3601 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     3661 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg
     3721 ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag
     3781 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcgctctcc acgcatcgtc
     3841 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgccctggc
     3901 ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg
     3961 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta
     4021 gctatagttc tagtggttgg ctacgtaccg gaatattaat agatcatgga gataattaaa
     4081 atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa
     4141 aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc ggatctcggt
     4201 ccgaaaccat gtcgtactac catcaccatc accatcacga ttacgatatc ccaacgggtg
     4261 gtaccatgta cgcaacgctt gaagaagcca ttgacgctgc acgcgaagaa tttcttgcag
     4321 acaaccccgg catcgacgcc gaagatgcga atgtgcaaca gttcaatgcc caaaaatacg
     4381 ttttgcagga cggcgacatc atgtggcaag ttgagttttt tgccgacgaa ggggaagaag
     4441 gtgaatgttt acctatgctt agcggtgaag ccgcgcaaag tgtttttgat ggcgactatg
     4501 atgagataga gatacgccag gagtggcagg aagagaatac attacatgaa tgggacgagg
     4561 gggaatttca gcttgagcca ccgctggata ccgaggaagg acgcgcagca gctgatgagt
     4621 gggatgaacg tggtacctct ctggttccgc gtggttcagg taccatgtac gcaacgcttg
     4681 aagaagccat tgacgctgca cgcgaagaat ttcttgcaga caaccccggc atcgacgccg
     4741 aagatgcgaa tgtgcaacag ttcaatgccc aaaaatacgt tttgcaggac ggcgacatca
     4801 tgtggcaagt tgagtttttt gccgacgaag gggaagaagg tgaatgttta cctatgctta
     4861 gcggtgaagc cgcgcaaagt gtttttgatg gcgactatga tgagatagag atacgccagg
     4921 agtggcagga agagaataca ttacatgaat gggacgaggg ggaatttcag cttgagccac
     4981 cgctggatac cgaggaagga cgcgcagcag ctgatgagtg ggatgaacgt ggtacctctc
     5041 tggttccgcg tggttcaggt accatgtacg caacgcttga agaagccatt gacgctgcac
     5101 gcgaagaatt tcttgcagac aaccccggca tcgacgccga agatgcgaat gtgcaacagt
     5161 tcaatgccca aaaatacgtt ttgcaggacg gcgacatcat gtggcaagtt gagttttttg
     5221 ccgacgaagg ggaagaaggt gaatgtttac ctatgcttag cggtgaagcc gcgcaaagtg
     5281 tttttgatgg cgactatgat gagatagaga tacgccagga gtggcaggaa gagaatacat
     5341 tacatgaatg ggacgagggg gaatttcagc ttgagccacc gctggatacc gaggaaggac
     5401 gcgcagcagc tgatgagtgg gatgaacgta gcagcggcct ggtgccgcgc ggcagccata
     5461 gcgaaaacct ctacttccaa tcgatggcgg aagaggagga ggtggaggag gggaggtcct
     5521 cgtcgtcggc gatactggac ctgccggagc cgctgctgct gcacatcctg agcttcctga
     5581 cggacgtgag gtctcggcac agggcggcgc tggcgtgcgg gaggatgcgg gcggcggagc
     5641 gggcgacgag gtcggagctc tcgctgaggg gcgacccgag gtcgccgggg ttcctgttcc
     5701 tctcgcacgc gttccgcttc ccggcgctgg aacacctcga cctctcgctc gtctcgccgt
     5761 gggggcatcc gcttctctcc tccgtgccgc cctgcggcgg cggcggcggc ggcgcgccct
     5821 cggcgtcgtc gtcgtcgggg atgaacgtgt accaccccga ggcgatctcc gagcagaacg
     5881 ccttcatcgc cgcccgcctc gcgggctgct tcccggcggt gacctcgctc gccgtctact
     5941 gccgcgaccc caccacgctc gccaacctca ccccgcactg gcaggcctcc ctccgccgcg
     6001 tcaagctcgt gcgctggcac cagcgcccgc ccaccctccc cgacggcgcg gatctcgagc
     6061 cgctgctgga gacctgcgcc gcgctccggg agctcgacct gtcggagttc tactgctgga
     6121 ccgaggacgt cgtgagggcg ctcaccacgc acccttccgc caccgcggcg ctcacccacc
     6181 tcgacctcgg cctcgccgcc gccaccgacg gcttcaaatc ctccgagctt gggccaatcg
     6241 cggcctcctg ccccaacctc cgcaagctcg tggcgccatg cttgttcaac ccacggttca
     6301 gcgattgcgt cggcgacgac gcgctgctct cgctggccac cagctgcccg cggctgaccg
     6361 tcttgcggct cagcgagccg ttcgaggctg cggccaacat ccagagggag gaggcggcca
     6421 tcaccgttgc ggggctagtc gccttcttcg cggcgctccc cgcgctggag gatttcacca
     6481 tggatctcca gcacaatgtg ctggaggccg cgcccgcgat ggaggcgctt gcccgaaggt
     6541 gcccgcggat caagttcttg accctgggtt ccttccaggg gctgtgtaag gcctcttggt
     6601 tgcatcttga tggtgttgcg gtgtgcggtg ggctggagtc actttacatg aagaattgcc
     6661 aggatctcac ggatgccagc cttgcggcaa ttggccgtgg gtgccggagg cttgctaagt
     6721 tcggcatcca tggctgtgac cttgtcactt cggctgggat caggaggctt gcattcacgc
     6781 ttcggcctac tctcaaggaa gtcactgtct tgcactgccg gcttctgcac actgcagaat
     6841 gtctcactgc tctaagtccg atccgtgatc gcattgaaag tcttgagatc aactgtgtct
     6901 ggaacacaac cgaaaacctg tacttccaat ctgaacaacc ctgcagtgtt gcaaatggca
     6961 ccaccaccga atgcgatcct gaggatgatg agcttggtga agtgtacgag tctgcagcca
     7021 agaaatgtag gtacatggaa tttgatgatg aaaacctgta cttccaatct cttggaagct
     7081 gggagatgct caggtcactc tccctatggt tctctgctgg ccagcttctc tctccgctca
     7141 tttctgctgg tctcgatagc tgtcccgtgc ttgaggagat ctcaattaag gtggagggtg
     7201 attgccggac atgcccacga cctgctccaa gaacaatttt tggcttaagt gatcttgcag
     7261 gcttcccagt attagccaag atgaaattgg acctcagtga agctgtgggt tatgcactta
     7321 ctgcaccaac agggcagatg gatctttcac tatgggagcg attttatttg catggtatcg
     7381 aatcactgca gactttgtat gaattggact actggccgcc ccaagacaag gatgtgcacc
     7441 accggagcct gacattgcca gccgtgggat tgatccaacg ctgcgttgga ctcaggaagc
     7501 ttttcatcca tggcaccaca catgagcact tcatgacctt cttcctttca attccaaact
     7561 tgcgggacat gcagttgcgg gaggactatt atccagcccc agagaatgat ctgatgttca
     7621 cagagatgcg ggctgaatct tggcttaggt ttgaggtgca actgaacagc cggcaaattg
     7681 atgattagcc ggaggatgag tgtggcctta agcaagcagc ggctcttgca gtctcgaggc
     7741 atgcggtacc aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt
     7801 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     7861 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     7921 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7981 caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagt aatcccacca
     8041 ctcctctcca cttaattaac ctagaggccg cgctggccgg aatattaata gatcatggag
     8101 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
     8161 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcgggaaaa
     8221 cctctacttc caatcgatgt ctgcgaagaa gattgtgttg aagagttccg atggtgaatc
     8281 tttcgaggtt gaggaggcgg tggctctcga gtcacaaacc atagcgcata tggttgaaga
     8341 cgactgcgtc gacaacggag tccctcttcc taacgtcacg agcaagatcc tcgccaaggt
     8401 gatcgagtat tgcaagaggc acgtcgaggc tgctgcctct aaggccgagg ccgtcgaggg
     8461 tgctgctacc tccgatgacg atcttaaggc ctgggacgct gattttatga agatcgatca
     8521 agctactctc tttgaactca ttctggctgc taattacctg aatatcaaga acttgcttga
     8581 tctaacatgt cagacagttg cggatatgat caaaggaaag actccagaag agatccgcac
     8641 aacgttcaac attaagaacg acttcacacc agaggaagag gaagaggttc gcagagagaa
     8701 ccaatgggct tttgaatgat aaccggagga tgagtgtggc ctgcagtctc gaggcatgcg
     8761 gtaccaagct tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg
     8821 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     8881 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     8941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     9001 tcatcaatgt atcttatcat gtctggatct gatcactgct tggccagggc ggcctaggag
     9061 atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     9121 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     9181 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac a
//
