LOCUS       sequence-478353-        4759 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  ScVip1-536-1107-E576A-K623A-Q625S-K627A protein expression in insect
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4759)
  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 4759)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 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-E576A-K623A-Q625S-K627A
FEATURES             Location/Qualifiers
     source          1..4759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  724..947
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1014..1547)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1736..1741,1742..1758,1759..1764))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1835..1868)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1945..2036
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1961..1978
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2081..2110
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(2120..2143,2144..2179,2180..2203)
                     /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             2219..2239
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4150..4284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4241..4260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4405..4570
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattgacttg ggtcgctctt cctgtggatg cgcagatgcc ctgcgtaagc
      721 gggtgtgggc ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa
      781 agtcttaaac tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag
      841 catactggac ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc
      901 gtcgtattaa agaggggcgt ggccaagggc atgtaaagac tatattcgcg gcgttgtgac
      961 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
     1021 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
     1081 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
     1141 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc
     1201 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
     1261 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
     1321 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
     1381 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
     1441 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
     1501 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
     1561 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     1621 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
     1681 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
     1741 acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg
     1801 tcacccggca accttgggca gcagcgaagt cgccataact tcgtatagca tacattatac
     1861 gaagttatct gtaactataa cggtcctaag gtagcgagtt taaacactag tatcgattcg
     1921 cgacctactc cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa
     1981 ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc
     2041 cggattattc ataccgtccc accatcgggc gcggatgaag catcatcatc atcatcacca
     2101 tcaccaccac ggtgctagct ggtcccaccc ccagttcgag aagggtggtg gttccggtgg
     2161 cggatccggt ggttctgctt ggagccaccc tcaattcgaa aagggctccg gttccggcga
     2221 gaacctgtac ttccaaggtg ccaaatgggt gttcaagggc ctcgccatca tcatccgtca
     2281 cgctgaccgt actcctaagc aaaagttcaa gcactccttc actagcccca tcttcatctc
     2341 cctgctgaag ggtcacaaag aggctgtcgt catccgtaac gtgaacgatc tgaagatcgt
     2401 gctgcaggct ctgcgtatcg ctctggacga gaaggctggc aaccccgcta agatcaaggt
     2461 gctggctaac gctctcgaga agaagctgaa cttccccggc accgctatct ccctggctcc
     2521 agtgctgaac aaagagaacg aggtcgagaa ggtgcagttc atcctgaagt ggggtggcga
     2581 gcctactcac tccgctaagt accaggctac cgagctggga gagcagatgc gtcaagactt
     2641 cgacctgttg aacaagtcca tcctgcagaa tatcaagatc ttcagctcct ccgagcgtcg
     2701 tgtgctgcac actgctcagt actggacccg tgctctgttc ggtgctgacg agctgggttc
     2761 cgacgagatc tctatccgca aggacctgct ggacgactcc aacgctgcta aggacctcat
     2821 ggacaaagtc aagaagaagt tgaagcccct gctgcgcgag ggcaaagagg ctcctcctca
     2881 attcgcttgg ccctccaaga tgcccgagcc ttacctggtc atcaagcgtg tggtcgagct
     2941 gatgaactac cacaagaaaa tcatggataa caacttcgcc aagaaagacg tgaactccat
     3001 gcagacccgt tggtgcacct ctgaggaccc cagcctgttc aaagagcgtt gggacaagct
     3061 tttcaaagag ttcaacaacg ccgagaaggt cgacccttcc aagatctccg agctgtacga
     3121 caccatgaag tacgacgctc tgcacaaccg tcagttcttg gagaacatct tcgaccccgg
     3181 cctgcctaac gaggctatcg ctgacgaact cggttctcac tccctggtgg atcgttaccc
     3241 catcaacgtc ctggccaaga acaacttcaa gatcatcgac tcccactcca tgaacaactc
     3301 cggcaagaac tcctccaact ccgtgggttc tctcggctgg gtgctcgagt ctggaaagac
     3361 ctcaaccgct cgtaacccca agtcctctag ccagttcgac gagccccgtt tcatgcagct
     3421 gcgcgaactg tacaagctgg ctaaggtcct gttcgacttc atctgcccca aagagtacgg
     3481 tatctccgac gctgagaagc tggacatcgg cctgctgact tctctgcctc tggctaagca
     3541 gatcctgaac gacatcggcg acatgaagaa ccgcgagact cccgcttgtg tggcttactt
     3601 caccaaagag tcccacatct acaccctgct caacatcatc tacgaaagcg gtatccccat
     3661 gaggatcgct cgcaacgctc tgcccgaact ggactacttg tcccagatca ccttcgagtt
     3721 gtacgagtcc accgacgctt ccggccagaa gtcccactct atccgtctga agatgtctcc
     3781 cggttgccac actcaggacc ctctggacgt gcaattggac gaccgtcact acatctcttg
     3841 catccctaag atctccctga ccaagcacct cgacatggac tacgtgcagc agaagctgcg
     3901 taacaagttc acccgtgtga tcatgcctcc aaagttcact cccgtgaaca tcacctcata
     3961 agcggccgca ctcgagtagg atcgaatcgg actttcgaat ctagagcctg cagtctcgac
     4021 aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta
     4081 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4141 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4201 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4261 aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctagaagatc cggctgctaa
     4321 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tatcataacc
     4381 cctagggtat acccatctaa ttggaaccag ataagtgaaa tctagttcca aactattttg
     4441 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
     4501 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
     4561 tccgcccaca accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt
     4621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4741 aaaagatcaa aggatcttc
//
