LOCUS       sequence-457780-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  AUTHORS   Wang S, Tsun ZY, Wolfson RL, Shen K, Wyant GA, Plovanich ME, Yuan 
            ED, Jones TD, Chantranupong L, Comb W, Wang T, Bar-Peled L, Zoncu R,
            Straub C, Kim C, Park J, Sabatini BL, Sabatini DM
  TITLE     Metabolism. Lysosomal amino acid transporter SLC38A9 signals 
            arginine sufficiency to mTORC1.
  JOURNAL   Science. 2015 Jan 9;347(6218):188-94. doi: 10.1126/science.1257132. 
            Epub 2015 Jan 7.
  PUBMED    25567906
REFERENCE   2  (bases 1 to 6404)
  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.1257132"; journalName: "Science"; date: "2015-01-9-
            9"; volume: "347"; issue: "6218"; pages: "188-94"
COMMENT     Sequence Label: pRK5-FLAG-SLC38A9.1
FEATURES             Location/Qualifiers
     source          1..6404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        92..449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        92..283
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(115..135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      300..435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(469..486)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(469..485)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(478..500)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      698..1153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(785..804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     995..1016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1170..1189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1289..1311
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1349..1367)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1435..1539
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1540..1608,1609..2400)
                     /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(1758..1777)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2571..3159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3060..3079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3313..3330
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3447..3468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3483..3488,3489..3506,3507..3513)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3521..3537
                     /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)."
     primer_bind     3526..3548
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3545..3561
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3545..3561
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3590..3969
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3970..4173
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4124..4144
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4170..4194
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        4404..4422
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4404..4421
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     4435..4454
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4513..4536
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6280..6289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    join(6293..6404,1..23)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6330..6349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6384..6403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 atgtatctta tcatgtctgg atcgggaatt aattcggcgc agcaccatgg cctgaaataa
       61 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
      121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      361 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
      421 gcttttttgg aggcctaggc ttttgcaaaa agctgttaac agcttggcac tggccgtcgt
      481 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      541 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      601 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
      661 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
      721 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      781 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      841 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      901 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      961 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1021 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     1081 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1141 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     1201 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     1261 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     1321 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     1381 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     1441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1501 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1561 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1621 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1681 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1741 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1801 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1861 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1921 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1981 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2041 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2101 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2161 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2221 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2281 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2341 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2401 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2701 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3181 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3241 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3301 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3361 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3421 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     3481 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     3541 cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg acattgatta
     3601 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     3661 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     3721 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     3781 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     3841 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     3901 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     3961 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4021 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4081 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4141 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     4201 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
     4261 ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc
     4321 tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa ttaatacata
     4381 accttatgta tcatacacat acgatttagg tgacactata gaataacatc cactttgcct
     4441 ttctctccac aggtgtccac tcccaggtcc aactgcacct cggttctatc gattgaattc
     4501 accatggaaa tggactacaa ggatgacgat gacaagggtg gtggtggtgg tgcgtcgaca
     4561 atggctaaca tgaacagtga ctcaagacat ctgggaacaa gcgaggtgga ccacgagcgg
     4621 gaccctggcc ctatgaacat tcagtttgag ccctccgacc tgcggtctaa gagacccttc
     4681 tgcatcgaac ctaccaacat tgtgaacgtc aatcacgtga tccagagggt cagtgatcat
     4741 gcatcagcca tgaataagcg aattcactac tattcccggc tgaccacacc cgctgacaaa
     4801 gcactgatcg cccctgatca tgtggtccca gcacccgagg aatgttacgt gtatagccct
     4861 ctgggatccg cctacaagct gcagagctac accgaggggt atggaaaaaa cacatccctg
     4921 gtgactattt ttatgatctg gaataccatg atggggacaa gtattctgtc aatcccatgg
     4981 ggaatcaagc aggccgggtt cactaccgga atgtgcgtga tcattctgat gggcctgctg
     5041 accctgtact gctgttatag ggtggtcaaa tctcgcacaa tgatgttttc actggacaca
     5101 actagctggg agtaccccga tgtgtgtcgg cactattttg gaagcttcgg ccagtggagc
     5161 tccctgctgt tcagcctggt gtccctgatt ggcgctatga tcgtgtactg ggtcctgatg
     5221 agcaacttcc tgtttaatac cgggaagttc atctttaact tcatccacca tatcaacgac
     5281 accgatacaa tcctgtctac aaacaatagt aaccctgtga tttgcccatc tgccggaagt
     5341 ggcgggcatc cagacaactc tagtatgatc ttttacgcta atgatactgg cgcacagcag
     5401 ttcgagaagt ggtgggacaa aagcagaacc gtgccctttt atctggtcgg cctgctgctg
     5461 cctctgctga actttaagtc cccatctttc tttagtaaat tcaatatcct ggggacagtg
     5521 tcagtcctgt acctgatttt tctggtgact ttcaaagcag tccgcctggg attccacctg
     5581 gaatttcatt ggttcatccc aactgagttc tttgtgcccg aaattcgatt tcagttccca
     5641 cagctgactg gagtgctgac cctggccttc tttatccaca attgcatcat tacactgctg
     5701 aagaacaata agaaacagga gaacaatgtg cgggacctgt gtatcgctta tatgctggtc
     5761 actctgacct acctgtatat tggggtgctg gtctttgcat cattccctag cccccctctg
     5821 tctaaagatt gtatcgaaca gaactttctg gacaatttcc catcaagcga tacactgagt
     5881 ttcattgcca gaatctttct gctgttccag atgatgactg tgtaccccct gctggggtat
     5941 ctggctaggg tccagctgct gggccacatt tttggggata tctaccctag cattttccat
     6001 gtgctgatcc tgaacctgat cattgtgggc gccggggtca tcatggcttg cttctaccct
     6061 aatattggag gcatcattag gtattccgga gccgcttgtg gcctggcttt tgtgttcatc
     6121 tacccatccc tgatctacat catttctctg caccaggagg aacgcctgac ctggcccaag
     6181 ctgatctttc atgtgttcat cattatcctg ggcgtggcaa acctgattgt ccagttcttt
     6241 atgtgagcgg ccgctaagta agtaaggatc cccagcttgg ccgccatggc ccaacttgtt
     6301 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6361 atttttttca ctgcattcta gttgtggttt gtccaaactc atca
//
