LOCUS       sequence-475909-        5989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ScVip1-536-1107-K817S-D819A-S821A protein expression in insect 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5989)
  AUTHORS   Raia P, Lee K, Bartsch SM, Rico-Resendiz F, Portugal-Calisto D, 
            Vadas O, Panse VG, Fiedler D, Hothorn M
  TITLE     A small signaling domain controls PPIP5K phosphatase activity in 
            phosphate homeostasis.
  JOURNAL   Nat Commun. 2025 Feb 19;16(1):1753. doi: 10.1038/s41467-025-56937-0.
  PUBMED    39966396
REFERENCE   2  (bases 1 to 5989)
  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: "2025-02-19"; volume: "16(1):1753. doi"; pages: " 
            10.1038/s41467-025-56937-0"
COMMENT     Sequence Label: pBB3-ScVip1-536-1107-K817S-D819A-S821A
FEATURES             Location/Qualifiers
     source          1..5989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..64)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(304..323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(762..781)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    806..854
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1109..1142
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(1213..1218,1219..1235,1236..1241)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1430..1963
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  2030..2253
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(2388..2976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2468..2487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  3166..3331
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    3452..3586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3476..3495)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3530..3549
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5497..5517)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(join(5533..5556,5557..5592,5593..5616))
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             complement(5626..5655)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(5700..5791)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(5758..5775)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        5836..5945
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             join(5987..5989,1..3,4..717)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
       61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      181 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
      241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      361 aaccgcatcg agctgaaggg tatcgacttc aaggaggacg gcaacatcct ggggcacaag
      421 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      481 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      601 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaggat
      721 cgaatcggag catgcggtac cgggagatgg gggaggctaa ctgaaacacg gaaggagaca
      781 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
      841 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
      901 accgagaccc cattgggacc aatacgcccg cgtttcttcc ttttccccac cccaaccccc
      961 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaagcc ctgccatagc
     1021 cactacgggt acgtttaaac cttccggagt aggtcgcgaa tcgatactag tgtttaaact
     1081 cgctacctta ggaccgttat agttacagat aacttcgtat aatgtatgct atacgaagtt
     1141 atggcgactt cgctgctgcc caaggttgcc gggtgacgca caccgtggaa acggatgaag
     1201 gcacgaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg
     1261 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     1321 gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag
     1381 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     1441 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat
     1501 gggcatcatt cgcacatgta ggctcggccc tgaccaagtc aaatccatgc gggctgctct
     1561 tgatcttttc ggtcgtgagt tcggagacgt agccacctac tcccaacatc agccggactc
     1621 cgattacctc gggaacttgc tccgtagtaa gacattcatc gcgcttgctg ccttcgacca
     1681 agaagcggtt gttggcgctc tcgcggctta cgttctgccc aagtttgagc agccgcgtag
     1741 tgagatctat atctatgatc tcgcagtctc cggcgagcac cggaggcagg gcattgccac
     1801 cgcgctcatc aatctcctca agcatgaggc caacgcgctt ggtgcttatg tgatctacgt
     1861 gcaagcagat tacggtgacg atcccgcagt ggctctctat acaaagttgg gcatacggga
     1921 agaagtgatg cactttgata tcgacccaag taccgccacc taacaattcg ttcaagccga
     1981 gatcggcttc ccggccgcgg agttgttcgg taaattgtca caacgccgcg aatatagtct
     2041 ttacatgccc ttggccacgc ccctctttaa tacgacgggc aatttgcact tcagaaaatg
     2101 aagagtttgc tttagccata acaaaagtcc agtatgcttt ttcacagcat aactggactg
     2161 atttcagttt acaactattc tgtctagttt aagactttat tgtcatagtt tagatctatt
     2221 ttgttcagtt taagacttta ttgtccgccc acacccgctt acgcagggca tctgcgcatc
     2281 cacaggaaga gcgacccaag tcaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2821 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3121 atatatgagt aaacttggtc tgacagttga cccaagtcaa ccggttgtgg gcggacaaaa
     3181 tagttgggaa ctgggagggg tggaaatgga gtttttaagg attatttagg gaagagtgac
     3241 aaaatagatg ggaactgggt gtagcgtcgt aagctaatac gaaaattaaa aatgacaaaa
     3301 tagtttggaa ctagatttca cttatctggt tccaattaga tgggtatacc ctaggggtta
     3361 tgatagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
     3421 agccggatct tctaggctca agcagtgatc agatccagac atgataagat acattgatga
     3481 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3541 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3601 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     3661 cctctacaaa tgtggtatgg ctgattatga tcctctagta cttctcgaca agcttgtcga
     3721 gactgcaggc tctagattcg aaagtccgat tcgatcctac tcgagtgcgg ccgcttatga
     3781 ggtgatgttc acgggagtga actttggagg catgatcaca cgggtgaact tgttacgcag
     3841 cttctgctgc acgtagtcca tgtcgaggtg cttggtcagg gagatcttag ggatgcaaga
     3901 gatgtagtga cggtcgtcca attgcacgtc cagagggtcc tgagtgtggc aaccgggaga
     3961 catcttcaga cggatagagt gggacttctg gccggaagcg tcggtggact cgtacaactc
     4021 gaaggtgatc tgggacaagt agtccagttc gggcagagcg ttgcgagcga tcctcatggg
     4081 gataccgctt tcgtagatga tgttgagcag ggtgtagatg tgggactctt tggtgaagta
     4141 agccacacaa gcgggagtct cgcggttctt catgtcgccg atgtcgttca ggatctgctt
     4201 agccagaggc agagaagtca gcaggccgat gtccagcttc tcagcgtcgg agataccgta
     4261 ctctttgggg cagatgaagt cgaacaggac cttagccagc ttgtacagtt cgcgcagctg
     4321 catgaaacgg ggctcgtcga actggctaga ggacttgggg ttacgagcgg ttgaggtctt
     4381 tccagactcg agcacccagc cgagagaacc cacggagttg gaggagttct tgccggagtt
     4441 gttcatggag tgggagtcga tgatcttgaa gttgttcttg gccaggacgt tgatggggta
     4501 acgatccacc agggagtgag aaccgagttc gtcagcgata gcctcgttag gcaggccggg
     4561 gtcgaagatg ttctccaaga actgacggtt gtgcagagcg tcgtacttca tggtgtcgta
     4621 cagctcggag atcttagcag gagcgacgga ctcggcgttg ttgaactctt tgaaaagctt
     4681 gtcccaacgc tctttgaaca ggctggggtc ctcagaggtg caccaacggg tctgcatgga
     4741 gttcacgtct ttcttggcga agttgttatc catgattttc ttgtggtagt tcatcagctc
     4801 gaccacacgc ttgatgacca ggtaaggctc gggcatcttg gagggccaag cgaattgagg
     4861 aggagcctct ttgccctcgc gcagcagggg cttcaacttc ttcttgactt tgtccatgag
     4921 gtccttagca gcgttggagt cgtccagcag gtccttgcgg atagagatct cgtcggaacc
     4981 cagctcgtca gcaccgaaca gagcacgggt ccagtactga gcagtgtgca gcacacgacg
     5041 ctcggaggag ctgaagatct tgatattctg caggatggac ttgttcaaca ggtcgaagtc
     5101 ttgacgcatc tgctctccca gctcggtagc ctggtactta gcggagtgag taggctcgcc
     5161 accccacttc aggatgaact gcaccttctc gacctcgttc tctttgttca gcactggctt
     5221 cagctggatc ttggtgccgg ggaagttcag cttcttctcg agagcgttag ccagcacctt
     5281 gatcttagcg gggttgccag ccttctcgtc cagagcgata cgcagagcct gcagcacgat
     5341 cttcagatcg ttcacgttac ggatgacgac ttcctctttg tgacccttca gcagggagat
     5401 gaagatgggg ctagtgaagg agtgcttgaa cttttgctta ggagtacggt cagcgtgacg
     5461 gatgatgatg gcgaggccct tgaacaccca tttggcacct tggaagtaca ggttctcgcc
     5521 ggaaccggag cccttttcga attgagggtg gctccaagca gaaccaccgg atccgccacc
     5581 ggaaccacca cccttctcga actgggggtg ggaccagcta gcaccgtggt ggtgatggtg
     5641 atgatgatga tgatgcttca tccgcgcccg atggtgggac ggtatgaata atccggaata
     5701 tttataggtt tttttattac aaaactgtta cgaaaacagt aaaatactta tttatttgcg
     5761 agatggttat cattttaatt atctccatga tctattaata ttccggagta ggtcgcgaat
     5821 cgatactagt atacggacct ttaattcaac ccaacacaat atattatagt taaataagaa
     5881 ttattatcaa atcatttgta tattaattaa aatactatac tgtaaattac attttattta
     5941 caatcactcg acgaagactt gatcacccgg gcggtccgaa accatgatg
//
