LOCUS       sequence-478355-        6411 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  ScVip1-536-1107-delta848-918-R547A-H548A-R551A protein expression in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6411)
  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 6411)
  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: 
            pFastBac1-ScVip1-536-1107-delta848-918-R547A-H548A-R551A
FEATURES             Location/Qualifiers
     source          1..6411
                     /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"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  894..1118
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1185..1718)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1907..1912,1913..1929,1930..1935))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2287..2378
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2303..2320
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2427..2456
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(2466..2489,2490..2525,2526..2549)
                     /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             2565..2585
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4284..4418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4321..4340)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4375..4394
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4447..4612
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4796..5250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4883..4902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5092..5113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5276..5380
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5381..5449,5450..6241)
                     /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(5599..5618)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
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 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa
      841 cctggcaaaa tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc
      901 ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac
      961 tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac
     1021 ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa
     1081 agaggggcgt ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg
     1141 aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt
     1201 gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact
     1261 gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc
     1321 tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag
     1381 actgcgagat catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa
     1441 gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta
     1501 ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg
     1561 tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg
     1621 ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga
     1681 ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa
     1741 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa
     1801 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     1861 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga
     1921 acaggcttat gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     1981 aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt
     2041 ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct
     2101 gtgcacggat ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc
     2161 ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg
     2221 tttgttcgcc caggactcta gctatagttc tagtggttgg ctacgtatac tccggaatat
     2281 taatagatca tggagataat taaaatgata accatctcgc aaataaataa gtattttact
     2341 gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc
     2401 ccaccatcgg gcgcggatcc atgaagcatc atcatcatca tcaccatcac caccacggtg
     2461 ctagctggtc ccacccccag ttcgagaagg gtggtggttc cggtggcgga tccggtggtt
     2521 ctgcttggag ccaccctcaa ttcgaaaagg gctccggttc cggcgagaac ctgtacttcc
     2581 aaggtgccaa atgggtgttc aagggcctcg ccatcatcat cgctgccgct gacgctactc
     2641 ctaagcaaaa gttcaagcac tccttcacta gccccatctt catctccctg ctgaagggtc
     2701 acaaagagga agtcgtcatc cgtaacgtga acgatctgaa gatcgtgctg caggctctgc
     2761 gtatcgctct ggacgagaag gctggcaacc ccgctaagat caaggtgctg gctaacgctc
     2821 tcgagaagaa gctgaacttc cccggcacca agatccagct gaagccagtg ctgaacaaag
     2881 agaacgaggt cgagaaggtg cagttcatcc tgaagtgggg tggcgagcct actcactccg
     2941 ctaagtacca ggctaccgag ctgggagagc agatgcgtca agacttcgac ctgttgaaca
     3001 agtccatcct gcagaatatc aagatcttca gctcctccga gcgtcgtgtg ctgcacactg
     3061 ctcagtactg gacccgtgct ctgttcggtg ctgacgagct gggttccgac gagatctcta
     3121 tccgcaagga cctgctggac gactccaacg ctgctaagga cctcatggac aaagtcaaga
     3181 agaagttgaa gcccctgctg cgcgagggca aagaggctcc tcctcaattc gcttggccct
     3241 ccaagatgcc cgagccttac ctggtcatca agcgtgtggt cgagctgatg aactaccaca
     3301 agaaaatcat ggataacaac ttcgccaaga aagacgtgaa ctccatgcag acccgttggt
     3361 gcacctctga ggaccccagc ctgttcaaag agcgttggga caagcttttc aaagagttca
     3421 acaacgccga gaaggtcgac ccttccaaga tctccgagct gtacgacacc atgaagtacg
     3481 acgctctgca caaccgtcag ttcttggaga acatcttcga ccccggctcc tctggtcagt
     3541 tcgacgagcc ccgtttcatg cagctgcgcg aactgtacaa gctggctaag gtcctgttcg
     3601 acttcatctg ccccaaagag tacggtatct ccgacgctga gaagctggac atcggcctgc
     3661 tgacttctct gcctctggct aagcagatcc tgaacgacat cggcgacatg aagaaccgcg
     3721 agactcccgc ttgtgtggct tacttcacca aagagtccca catctacacc ctgctcaaca
     3781 tcatctacga aagcggtatc cccatgagga tcgctcgcaa cgctctgccc gaactggact
     3841 acttgtccca gatcaccttc gagttgtacg agtccaccga cgcttccggc cagaagtccc
     3901 actctatccg tctgaagatg tctcccggtt gccacactca ggaccctctg gacgtgcaat
     3961 tggacgaccg tcactacatc tcttgcatcc ctaagatctc cctgaccaag cacctcgaca
     4021 tggactacgt gcagcagaag ctgcgtaaca agttcacccg tgtgatcatg cctccaaagt
     4081 tcactcccgt gaacatcacc tcataagcgg ccgctttcga atctagagcc tgcagtctcg
     4141 aggcatgcgg taccaagctt gtcgagaagt actagaggat cataatcagc cataccacat
     4201 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     4261 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4321 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4381 tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagga
     4441 gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     4501 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     4561 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     4621 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     4681 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     4741 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga atgggacgcg
     4801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4981 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct
     5101 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5161 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5221 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     5281 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5341 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5401 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5461 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5521 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5581 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5641 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5701 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5761 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5821 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5881 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5941 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6001 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6061 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6121 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6181 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6241 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6301 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6361 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
