LOCUS       sequence-475917-        5989 bp    DNA     circular SYN 12-JUN-2025
DEFINITION  ScVip1-536-1107-K554A 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 Jun 12, 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-K554A
FEATURES             Location/Qualifiers
     source          1..5989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1478..1498)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(join(1514..1537,1538..1573,1574..1597))
                     /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(1607..1636)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        complement(1681..1772)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1739..1756)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        1817..1926
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             join(1968..1970,1971..1973,1974..2687)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2013..2034)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2274..2293)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2621..2642
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2732..2751)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2776..2824
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    3079..3112
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(3183..3188,3189..3205,3206..3211)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3400..3933
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4000..4223
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(4358..4946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4438..4457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  5136..5301
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    5422..5556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5465)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5500..5519
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 cgaaggtgat ctgggacaag tagtccagtt cgggcagagc gttgcgagcg atcctcatgg
       61 ggataccgct ttcgtagatg atgttgagca gggtgtagat gtgggactct ttggtgaagt
      121 aagccacaca agcgggagtc tcgcggttct tcatgtcgcc gatgtcgttc aggatctgct
      181 tagccagagg cagagaagtc agcaggccga tgtccagctt ctcagcgtcg gagataccgt
      241 actctttggg gcagatgaag tcgaacagga ccttagccag cttgtacagt tcgcgcagct
      301 gcatgaaacg gggctcgtcg aactggctag aggacttggg gttacgagcg gttgaggtct
      361 ttccagactc gagcacccag ccgagagaac ccacggagtt ggaggagttc ttgccggagt
      421 tgttcatgga gtgggagtcg atgatcttga agttgttctt ggccaggacg ttgatggggt
      481 aacgatccac cagggagtga gaaccgagtt cgtcagcgat agcctcgtta ggcaggccgg
      541 ggtcgaagat gttctccaag aactgacggt tgtgcagagc gtcgtacttc atggtgtcgt
      601 acagctcgga gatcttggaa gggtcgacct tctcggcgtt gttgaactct ttgaaaagct
      661 tgtcccaacg ctctttgaac aggctggggt cctcagaggt gcaccaacgg gtctgcatgg
      721 agttcacgtc tttcttggcg aagttgttat ccatgatttt cttgtggtag ttcatcagct
      781 cgaccacacg cttgatgacc aggtaaggct cgggcatctt ggagggccaa gcgaattgag
      841 gaggagcctc tttgccctcg cgcagcaggg gcttcaactt cttcttgact ttgtccatga
      901 ggtccttagc agcgttggag tcgtccagca ggtccttgcg gatagagatc tcgtcggaac
      961 ccagctcgtc agcaccgaac agagcacggg tccagtactg agcagtgtgc agcacacgac
     1021 gctcggagga gctgaagatc ttgatattct gcaggatgga cttgttcaac aggtcgaagt
     1081 cttgacgcat ctgctctccc agctcggtag cctggtactt agcggagtga gtaggctcgc
     1141 caccccactt caggatgaac tgcaccttct cgacctcgtt ctctttgttc agcactggct
     1201 tcagctggat cttggtgccg gggaagttca gcttcttctc gagagcgtta gccagcacct
     1261 tgatcttagc ggggttgcca gccttctcgt ccagagcgat acgcagagcc tgcagcacga
     1321 tcttcagatc gttcacgtta cggatgacga cttcctcttt gtgacccttc agcagggaga
     1381 tgaagatggg gctagtgaag gagtgcttga acttttgcgc aggagtacgg tcagcgtgac
     1441 ggatgatgat ggcgaggccc ttgaacaccc atttggcacc ttggaagtac aggttctcgc
     1501 cggaaccgga gcccttttcg aattgagggt ggctccaagc agaaccaccg gatccgccac
     1561 cggaaccacc acccttctcg aactgggggt gggaccagct agcaccgtgg tggtgatggt
     1621 gatgatgatg atgatgcttc atccgcgccc gatggtggga cggtatgaat aatccggaat
     1681 atttataggt ttttttatta caaaactgtt acgaaaacag taaaatactt atttatttgc
     1741 gagatggtta tcattttaat tatctccatg atctattaat attccggagt aggtcgcgaa
     1801 tcgatactag tatacggacc tttaattcaa cccaacacaa tatattatag ttaaataaga
     1861 attattatca aatcatttgt atattaatta aaatactata ctgtaaatta cattttattt
     1921 acaatcactc gacgaagact tgatcacccg ggcggtccga aaccatgatg gtgagcaagg
     1981 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2041 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2101 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     2161 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     2221 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2281 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2341 agctgaaggg tatcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2401 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2461 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2521 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2581 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2641 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaggat cgaatcggag
     2701 catgcggtac cgggagatgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     2761 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     2821 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     2881 cattgggacc aatacgcccg cgtttcttcc ttttccccac cccaaccccc aagttcgggt
     2941 gaaggcccag ggctcgcagc caacgtcggg gcggcaagcc ctgccatagc cactacgggt
     3001 acgtttaaac cttccggagt aggtcgcgaa tcgatactag tgtttaaact cgctacctta
     3061 ggaccgttat agttacagat aacttcgtat aatgtatgct atacgaagtt atggcgactt
     3121 cgctgctgcc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3181 agttgacata agcctgttcg gttcgtaaac tgtaatgcaa gtagcgtatg cgctcacgca
     3241 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3301 cttgttatga ctgttttttt gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     3361 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagcaacgat
     3421 gttacgcagc agggcagtcg ccctaaaaca aagttaggtg gctcaagtat gggcatcatt
     3481 cgcacatgta ggctcggccc tgaccaagtc aaatccatgc gggctgctct tgatcttttc
     3541 ggtcgtgagt tcggagacgt agccacctac tcccaacatc agccggactc cgattacctc
     3601 gggaacttgc tccgtagtaa gacattcatc gcgcttgctg ccttcgacca agaagcggtt
     3661 gttggcgctc tcgcggctta cgttctgccc aagtttgagc agccgcgtag tgagatctat
     3721 atctatgatc tcgcagtctc cggcgagcac cggaggcagg gcattgccac cgcgctcatc
     3781 aatctcctca agcatgaggc caacgcgctt ggtgcttatg tgatctacgt gcaagcagat
     3841 tacggtgacg atcccgcagt ggctctctat acaaagttgg gcatacggga agaagtgatg
     3901 cactttgata tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc
     3961 ccggccgcgg agttgttcgg taaattgtca caacgccgcg aatatagtct ttacatgccc
     4021 ttggccacgc ccctctttaa tacgacgggc aatttgcact tcagaaaatg aagagtttgc
     4081 tttagccata acaaaagtcc agtatgcttt ttcacagcat aactggactg atttcagttt
     4141 acaactattc tgtctagttt aagactttat tgtcatagtt tagatctatt ttgttcagtt
     4201 taagacttta ttgtccgccc acacccgctt acgcagggca tctgcgcatc cacaggaaga
     4261 gcgacccaag tcaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4321 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4381 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4441 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4501 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4561 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4621 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4681 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4741 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4801 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4861 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4921 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4981 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5041 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5101 aaacttggtc tgacagttga cccaagtcaa ccggttgtgg gcggacaaaa tagttgggaa
     5161 ctgggagggg tggaaatgga gtttttaagg attatttagg gaagagtgac aaaatagatg
     5221 ggaactgggt gtagcgtcgt aagctaatac gaaaattaaa aatgacaaaa tagtttggaa
     5281 ctagatttca cttatctggt tccaattaga tgggtatacc ctaggggtta tgatagttat
     5341 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct
     5401 tctaggctca agcagtgatc agatccagac atgataagat acattgatga gtttggacaa
     5461 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     5521 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     5581 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     5641 tgtggtatgg ctgattatga tcctctagta cttctcgaca agcttgtcga gactgcaggc
     5701 tctagattcg aaagtccgat tcgatcctac tcgagtgcgg ccgcttatga ggtgatgttc
     5761 acgggagtga actttggagg catgatcaca cgggtgaact tgttacgcag cttctgctgc
     5821 acgtagtcca tgtcgaggtg cttggtcagg gagatcttag ggatgcaaga gatgtagtga
     5881 cggtcgtcca attgcacgtc cagagggtcc tgagtgtggc aaccgggaga catcttcaga
     5941 cggatagagt gggacttctg gccggaagcg tcggtggact cgtacaact
//
