LOCUS       sequence-371455-        9275 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of SLC38A9 N-terminus and FNIP2 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9275)
  AUTHORS   Jansen RM, Peruzzo R, Fromm SA, Yokom AL, Zoncu R, Hurley JH
  TITLE     Structural basis for FLCN RagC GAP activation in MiT-TFE 
            substrate-selective mTORC1 regulation.
  JOURNAL   Sci Adv. 2022 Sep 16;8(37):eadd2926. doi: 10.1126/sciadv.add2926. 
            Epub 2022 Sep 14.
  PUBMED    36103527
REFERENCE   2  (bases 1 to 9275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2022-09-16"; volume: "8(37):eadd2926. doi"; pages: " 
            10.1126/sciadv.add2926. Epub 2022 Sep 14"
COMMENT     Sequence Label: pCAG-GST-SLC-FNIP2
FEATURES             Location/Qualifiers
     source          1..9275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1753..1762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1759..2415
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2373..2395
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2443..2463
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(6297..6316)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6362..6417
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6416..6435)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6778..6794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6778..6794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6791..6813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6802..6818
                     /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(6826..6832,6833..6850,6851..6856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6871..6892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6951..7146
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6997..7132
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7152..7286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7189..7208)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7243..7262
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7354..7371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7525..8113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7605..7624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8284..9075,9076..9144))
                     /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     8907..8926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9145..9249)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgcaggggg gggggggggg gggggggggg
      541 gggggggggg ggggcggrga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgatc
     1741 gacggtatcg atgccaccat ggcatcccct atactaggtt attggaaaat taagggcctt
     1801 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
     1861 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
     1921 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
     1981 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
     2041 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
     2101 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     2161 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     2221 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     2281 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     2341 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     2401 gaccatcctc caaaatcgga tgattacgat atcccaacga ccgaaaacct gtattttcag
     2461 ggtggtacca tggctaacat gaacagtgac tcaagacatc tgggaacaag cgaggtggac
     2521 cacgagcggg accctggccc tatgaacatt cagtttgagc cctccgacct gcggtctaag
     2581 agacccttct gcatcgaacc taccaacatt gtgaacgtca atcacgtgat ccagagggtc
     2641 agtgatcatg catcagccat gaataagcga attcactact attcccggct gaccacaccc
     2701 gctgacaaag cactgatcgc ccctgatcat gtggtcccag cacccgagga atgttacgtg
     2761 tatagccctc tgggatccgc ctacaagctg cagagctaca ccgaggggta tggaaaaaac
     2821 acatccgggt ctggctccgg atcagggagc ggctccggta ccatggcccc caccctgctg
     2881 cagaagctgt ttaataagag aggcagctcc ggcagcagcg ccgccgccag cgcccagggc
     2941 agggccccta aggagggccc agccttttcc tggtcttgca gcgagttcga cctgaacgag
     3001 atccgcctga tcgtgtacca ggactgtgat cggagaggcc ggcaggtgct gttcgacagc
     3061 aaggccgtgc agaagatcga ggaggtgacc gcccagaaga cagaggatgt gcctatcaag
     3121 atctccgcca agtgctgtca gggctcctct agcgtgtcct ctagctcctc tagctccatc
     3181 tctagccact cctctagcgg aggctcctct caccacgcaa aggagcagct gcccaagtac
     3241 cagtatacaa gacctgccag cgatgtgaac atgctgggcg agatgatgtt tggctctgtg
     3301 gccatgagct acaagggctc caccctgaag atccactata tccgctcccc ccctcagctg
     3361 atgatctcta aggtgtttag cgcccggatg ggctccttct gcggctctac aaacaatctg
     3421 caggacagct tcgagtacat caatcaggac cccaacctgg gcaagctgaa caccaatcag
     3481 aactccctgg gcccttgtag gaccggctcc aatctggccc actctacacc cgtggacatg
     3541 ccttctaggg gccagaacga ggacagggat agcggaatcg cccggtccgc ctctctgagc
     3601 tccctgctga tcacaccatt tcccagccct tctagctcca cctctagctc ctctagctat
     3661 cagaggcgct ggctgagaag ccagaccaca tccctggaga atggcatcat cccccggaga
     3721 tctaccgacg agacattcag cctggccgag gagacatgct cctctaaccc tgctatggtg
     3781 cggcgcaaga agatcgccat ctctatcatc tttagcctgt gcgagaagga ggaggcccag
     3841 aggaatttcc aggatttctt tttcagccac tttcccctgt tcgagtccca catgaacaga
     3901 ctgaagtctg ccatcgagaa ggccatgatc tcctgcagga agatcgccga gagctccctg
     3961 agagtgcagt tttacgtgtc taggctgatg gaggccctgg gcgagttcag aggcaccatc
     4021 tggaatctgt attctgtgcc aaggatcgcc gagcccgtgt ggctgaccat gatgagcggc
     4081 acactggaga agaaccagct gtgccagcgg ttcctgaagg agttcacact gctgatcgag
     4141 cagatcaata agaaccagtt tttcgccgcc ctgctgaccg ccgtgctgac ataccacctg
     4201 gcctgggtgc caaccgtgat gcccgtggac cacccaccca tcaaggcctt tagcgagaag
     4261 cggacaagcc agtccgtgaa tatgctggcc aagacccacc catacaaccc actgtgggca
     4321 cagctgggcg acctgtatgg agcaatcggc tcccctgtgc ggctgaccag aacagtggtg
     4381 gtgggcaagc agaaggatct ggtgcagcgc atcctgtacg tgctgacata tttcctgcgg
     4441 tgcagcgagc tgcaggagaa tcagctgacc tggtccggaa accacggcga gggcgatcag
     4501 gtgctgaatg gctccaagat catcaccgcc ctggagaagg gcgaggtgga ggagtctgag
     4561 tatgtggtca tcacagtgag aaatgagcca gccctggtgc ctccaatcct gccacctacc
     4621 gcagcagaga ggcacaaccc ctggcccaca ggctttcctg agtgcccaga gggcaccgac
     4681 agccgcgatc tgggcctgaa gccagacaag gaggccaaca gacggcccga gcagggctcc
     4741 gaggcctgct ctgccggatg tctgggacct gcctccgatg cctcttggaa gccacagaat
     4801 gccttctgtg gcgacgagaa gaacaaggag gcaccacagg atggctctag ccggctgcct
     4861 tcctgcgagg tgctgggagc cggcatgaag atggaccagc aggccgtgtg cgagctgctg
     4921 aaggtggaga tgcctacaag gctgccagac cgcagcgtgg catggccttg cccagatagg
     4981 cacctgcgcg agaagccttc cctggagaag gtgacctttc agatcggcag cttcgcctcc
     5041 ccagagtctg attttgagag ccgcatgaag aagatggagg agcgggtgaa ggcctgtggc
     5101 ccaagcctgg aggcctccga ggcagcagac gtggcacagg acccccaggt gtctcgcagc
     5161 ccctttaagc ctggcttcca ggagaacgtg tgctgtccac agaaccggct gagcgagggc
     5221 gacgagggag agtccgataa gggcttcgcc gaggaccggg gcagcagaaa cgacatggcc
     5281 gccgatatcg ccggccagct gtcccacgcc gccgatctgg gcaccgcctc tcacggcgca
     5341 ggaggcacag gcggcaggcg cctggaggca acccggggcc tgtacgtgaa ggcagccgag
     5401 ggaccagtgc tggagccagt ggcaccacgg tgcgtgcaga gaggaccagg cctggtggca
     5461 ggagcaaata tcccctgtgg cgacgataat aagaaggcca actttagaac cgagggcgac
     5521 atccccagga acgagtcctc tgacagcgcc ctgggcgatt ccgacgatga ggcctgcgcc
     5581 tctgccatgc tggacctggg ccacggcggc gatcgcacag gaggcagcct ggaggtggag
     5641 ctgccactgc ctcggtccca gtctatcagc acccagaatg tgaggaactt cggccgcagc
     5701 ctgctggccg gatactgccc cacatatatg cctgacctgg tgctgcacgg caccggctcc
     5761 gatgagaagc tgaagcagtg tctggtggcc gacctggtgc acacagtgca ccaccctgtg
     5821 ctggatgagc caatcgccga ggccgtgtgc atcatcgccg acaccgataa gtggtccgtg
     5881 caggtggcca cctctcagag aaaggtgaca gacaatatga agctgggcca ggatgtgctg
     5941 gtgagctccc aggtgtctag cctgctgcag agcatcctgc agctgtacaa gctgcacctg
     6001 cccgccgact tctgtatcat gcacctggag gataggctgc aggagatgta cctgaagtct
     6061 aagatgctga gcgagtatct gcggggccac accagagtgc acgtgaagga gctgggcgtg
     6121 gtgctgggca tcgagtccaa cgacctgcct ctgctgaccg ccatcgccag cacacactcc
     6181 ccatatgtgg cccagatcct gctgtaactc gaggaattca ctcctcaggt gcaggctgcc
     6241 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     6301 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     6361 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     6421 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     6481 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     6541 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     6601 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     6661 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     6721 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     6781 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6841 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6901 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     6961 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7021 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7081 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     7141 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7201 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7261 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     7321 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7381 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7441 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7501 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7561 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7621 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7681 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7741 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7801 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7861 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7921 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7981 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8041 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8101 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8161 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8221 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8281 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8341 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8401 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8461 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8521 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8581 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8641 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8701 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8761 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8821 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8881 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8941 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     9001 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9061 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9121 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9181 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9241 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
