LOCUS       sequence-338365-        8828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FNIP2 expressing vector for mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8828)
  AUTHORS   Lawrence RE, Fromm SA, Fu Y, Yokom AL, Kim DJ, Thelen AM, Young LN, 
            Lim CY, Samelson AJ, Hurley JH, Zoncu R
  TITLE     Structural mechanism of a Rag GTPase activation checkpoint by the 
            lysosomal folliculin complex.
  JOURNAL   Science. 2019 Nov 22;366(6468):971-977. doi: 
            10.1126/science.aax0364. Epub 2019 Oct 31.
  PUBMED    31672913
REFERENCE   2  (bases 1 to 8828)
  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: 
            "10.1126/science.aax0364"; journalName: "Science"; date: 
            "2019-11-22- 22"; volume: "366"; issue: "6468"; pages: "971-977"
COMMENT     Sequence Label: pCAG-GST FNIP2
FEATURES             Location/Qualifiers
     source          1..8828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          112..1128
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1052..1074
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1136..1155
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1202..1211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1208..1864
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1822..1844
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1892..1912
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5353..5372)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5418..5473
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5472..5491)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5834..5850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5834..5850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5847..5869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5858..5874
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5882..5888,5889..5906,5907..5912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5927..5948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6007..6202
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6053..6188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6208..6342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6245..6264)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6299..6318
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6410..6427)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6581..7169)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6661..6680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7340..8131,8132..8200))
                     /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     7963..7982
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8201..8305)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8336..8715
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
       61 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      121 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      181 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      241 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
      301 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
      361 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
      421 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
      481 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
      541 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
      601 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
      661 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
      721 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
      781 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
      841 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
      901 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
      961 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     1021 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     1081 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     1141 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgatcg acggtatcga
     1201 tgccaccatg gcatccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     1261 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     1321 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     1381 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     1441 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     1501 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     1561 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     1621 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     1681 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     1741 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     1801 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     1861 aaaatcggat gattacgata tcccaacgac cgaaaacctg tattttcagg gtggtaccat
     1921 ggcccccacc ctgctgcaga agctgtttaa taagagaggc agctccggca gcagcgccgc
     1981 cgccagcgcc cagggcaggg cccctaagga gggcccagcc ttttcctggt cttgcagcga
     2041 gttcgacctg aacgagatcc gcctgatcgt gtaccaggac tgtgatcgga gaggccggca
     2101 ggtgctgttc gacagcaagg ccgtgcagaa gatcgaggag gtgaccgccc agaagacaga
     2161 ggatgtgcct atcaagatct ccgccaagtg ctgtcagggc tcctctagcg tgtcctctag
     2221 ctcctctagc tccatctcta gccactcctc tagcggaggc tcctctcacc acgcaaagga
     2281 gcagctgccc aagtaccagt atacaagacc tgccagcgat gtgaacatgc tgggcgagat
     2341 gatgtttggc tctgtggcca tgagctacaa gggctccacc ctgaagatcc actatatccg
     2401 ctccccccct cagctgatga tctctaaggt gtttagcgcc cggatgggct ccttctgcgg
     2461 ctctacaaac aatctgcagg acagcttcga gtacatcaat caggacccca acctgggcaa
     2521 gctgaacacc aatcagaact ccctgggccc ttgtaggacc ggctccaatc tggcccactc
     2581 tacacccgtg gacatgcctt ctaggggcca gaacgaggac agggatagcg gaatcgcccg
     2641 gtccgcctct ctgagctccc tgctgatcac accatttccc agcccttcta gctccacctc
     2701 tagctcctct agctatcaga ggcgctggct gagaagccag accacatccc tggagaatgg
     2761 catcatcccc cggagatcta ccgacgagac attcagcctg gccgaggaga catgctcctc
     2821 taaccctgct atggtgcggc gcaagaagat cgccatctct atcatcttta gcctgtgcga
     2881 gaaggaggag gcccagagga atttccagga tttctttttc agccactttc ccctgttcga
     2941 gtcccacatg aacagactga agtctgccat cgagaaggcc atgatctcct gcaggaagat
     3001 cgccgagagc tccctgagag tgcagtttta cgtgtctagg ctgatggagg ccctgggcga
     3061 gttcagaggc accatctgga atctgtattc tgtgccaagg atcgccgagc ccgtgtggct
     3121 gaccatgatg agcggcacac tggagaagaa ccagctgtgc cagcggttcc tgaaggagtt
     3181 cacactgctg atcgagcaga tcaataagaa ccagtttttc gccgccctgc tgaccgccgt
     3241 gctgacatac cacctggcct gggtgccaac cgtgatgccc gtggaccacc cacccatcaa
     3301 ggcctttagc gagaagcgga caagccagtc cgtgaatatg ctggccaaga cccacccata
     3361 caacccactg tgggcacagc tgggcgacct gtatggagca atcggctccc ctgtgcggct
     3421 gaccagaaca gtggtggtgg gcaagcagaa ggatctggtg cagcgcatcc tgtacgtgct
     3481 gacatatttc ctgcggtgca gcgagctgca ggagaatcag ctgacctggt ccggaaacca
     3541 cggcgagggc gatcaggtgc tgaatggctc caagatcatc accgccctgg agaagggcga
     3601 ggtggaggag tctgagtatg tggtcatcac agtgagaaat gagccagccc tggtgcctcc
     3661 aatcctgcca cctaccgcag cagagaggca caacccctgg cccacaggct ttcctgagtg
     3721 cccagagggc accgacagcc gcgatctggg cctgaagcca gacaaggagg ccaacagacg
     3781 gcccgagcag ggctccgagg cctgctctgc cggatgtctg ggacctgcct ccgatgcctc
     3841 ttggaagcca cagaatgcct tctgtggcga cgagaagaac aaggaggcac cacaggatgg
     3901 ctctagccgg ctgccttcct gcgaggtgct gggagccggc atgaagatgg accagcaggc
     3961 cgtgtgcgag ctgctgaagg tggagatgcc tacaaggctg ccagaccgca gcgtggcatg
     4021 gccttgccca gataggcacc tgcgcgagaa gccttccctg gagaaggtga cctttcagat
     4081 cggcagcttc gcctccccag agtctgattt tgagagccgc atgaagaaga tggaggagcg
     4141 ggtgaaggcc tgtggcccaa gcctggaggc ctccgaggca gcagacgtgg cacaggaccc
     4201 ccaggtgtct cgcagcccct ttaagcctgg cttccaggag aacgtgtgct gtccacagaa
     4261 ccggctgagc gagggcgacg agggagagtc cgataagggc ttcgccgagg accggggcag
     4321 cagaaacgac atggccgccg atatcgccgg ccagctgtcc cacgccgccg atctgggcac
     4381 cgcctctcac ggcgcaggag gcacaggcgg caggcgcctg gaggcaaccc ggggcctgta
     4441 cgtgaaggca gccgagggac cagtgctgga gccagtggca ccacggtgcg tgcagagagg
     4501 accaggcctg gtggcaggag caaatatccc ctgtggcgac gataataaga aggccaactt
     4561 tagaaccgag ggcgacatcc ccaggaacga gtcctctgac agcgccctgg gcgattccga
     4621 cgatgaggcc tgcgcctctg ccatgctgga cctgggccac ggcggcgatc gcacaggagg
     4681 cagcctggag gtggagctgc cactgcctcg gtcccagtct atcagcaccc agaatgtgag
     4741 gaacttcggc cgcagcctgc tggccggata ctgccccaca tatatgcctg acctggtgct
     4801 gcacggcacc ggctccgatg agaagctgaa gcagtgtctg gtggccgacc tggtgcacac
     4861 agtgcaccac cctgtgctgg atgagccaat cgccgaggcc gtgtgcatca tcgccgacac
     4921 cgataagtgg tccgtgcagg tggccacctc tcagagaaag gtgacagaca atatgaagct
     4981 gggccaggat gtgctggtga gctcccaggt gtctagcctg ctgcagagca tcctgcagct
     5041 gtacaagctg cacctgcccg ccgacttctg tatcatgcac ctggaggata ggctgcagga
     5101 gatgtacctg aagtctaaga tgctgagcga gtatctgcgg ggccacacca gagtgcacgt
     5161 gaaggagctg ggcgtggtgc tgggcatcga gtccaacgac ctgcctctgc tgaccgccat
     5221 cgccagcaca cactccccat atgtggccca gatcctgctg taactcgagg aattcactcc
     5281 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     5341 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     5401 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     5461 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     5521 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     5581 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     5641 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     5701 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     5761 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     5821 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5881 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5941 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6001 ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6181 tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa tggttacaaa
     6241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6301 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat taatgaatcg
     6361 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6421 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6481 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6541 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6601 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6661 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6721 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6781 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6841 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6901 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6961 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7021 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7081 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7141 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7201 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7261 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7321 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7381 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7441 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7501 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7561 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7621 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7681 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7741 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7801 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7861 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7921 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7981 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8041 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8101 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8161 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8221 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8281 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     8341 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     8401 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     8461 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     8521 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     8581 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     8641 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     8701 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     8761 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     8821 gcgggggg
//
