LOCUS       sequence-314411-        5788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Untagged SEH1L cDNA expression vector for transfections.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5788)
  AUTHORS   Orozco JM, Krawczyk PA, Scaria SM, Cangelosi AL, Chan SH, Kunchok T,
            Lewis CA, Sabatini DM
  TITLE     Dihydroxyacetone phosphate signals glucose availability to mTORC1.
  JOURNAL   Nat Metab. 2020 Sep;2(9):893-901. doi: 10.1038/s42255-020-0250-5. 
            Epub 2020 Jul 27.
  PUBMED    32719541
REFERENCE   2  (bases 1 to 5788)
  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: 
            "10.1038/s42255-020-0250-5"; journalName: "Nat Metab"; date: 
            "2020-09"; volume: "2"; issue: "9"; pages: "893-901"
COMMENT     Sequence Label: SEH1L in pRK5
FEATURES             Location/Qualifiers
     source          1..5788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2028..2054
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2075..2084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2088..2222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2125..2144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2179..2198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2291..2648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2291..2482
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2314..2334)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2499..2634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2668..2685)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2668..2684)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2677..2699)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2897..3352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2984..3003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3194..3215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3369..3388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3488..3510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3548..3566)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3634..3738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3739..3807,3808..4599)
                     /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(3957..3976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4770..5358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5259..5278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5512..5529
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5646..5667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5682..5687,5688..5705,5706..5712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5720..5736
                     /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     5725..5747
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5744..5760
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5744..5760
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgattgaatt cgtcgacgcc accatgtttg tggctcgcag catcgcggcg gaccacaagg
      961 atctcatcca cgatgtctct ttcgacttcc acgggcggcg gatggcaacc tgctccagcg
     1021 atcagagcgt taaggtctgg gataaaagtg aaagtggtga ttggcattgt actgctagct
     1081 ggaagacaca tagtggatct gtatggcgtg tgacatgggc ccatcctgaa tttgggcagg
     1141 ttttggcttc ctgttctttt gaccgaacag ctgctgtatg ggaagaaata gtaggagaat
     1201 caaatgataa actgcgagga cagagccact gggttaaaag gacaactctg gtggatagca
     1261 gaacatctgt tactgatgtg aagtttgctc ccaagcacat gggtcttatg ttagcaacct
     1321 gttccgcaga tggtatagta agaatctatg aggcaccaga tgttatgaat ctcagccagt
     1381 ggtctttgca gcatgagatc tcatgtaagc taagctgtag ttgtatttct tggaaccctt
     1441 caagctctcg tgctcattcc cccatgatcg ccgtaggaag tgatgacagt agccccaacg
     1501 caatggccaa ggttcagatt tttgaatata atgaaaacac caggaaatat gcaaaagctg
     1561 aaactcttat gacagtcact gatcctgttc atgatattgc attcgctcca aatttgggaa
     1621 gatctttcca tattctagca atagcgacca aagatgtgag aatttttaca ttaaagcctg
     1681 tgaggaaaga actgacttcc tctggtgggc caacaaagtt tgaaatccat atagtggctc
     1741 agttcgataa tcataattct caggtctggc gagtgagttg gaatataaca ggaacggtgc
     1801 tagcatcttc aggagatgat gggtgtgtaa gattgtggaa agctaattat atggacaatt
     1861 ggaagtgtac tggtattttg aaaggtaatg ggagcccagt caatgggagt tctcagcagg
     1921 gaacctcaaa tccttcccta ggttcaacta ttccaagtct tcagaattca ttaaatggat
     1981 cttctgctgg cagaaagcac agctgagcgg ccgcaggagg tggaggttac ccctatgatg
     2041 tgcctgacta cgcatgataa ggatccccag cttggccgcc atggcccaac ttgtttattg
     2101 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2161 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     2221 tcgggaatta attcggcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     2281 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2401 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2461 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2521 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2581 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2641 tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     2701 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     2761 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2821 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     2881 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     3241 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3301 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     3361 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3421 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3481 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3541 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3601 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3661 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3721 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3781 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3841 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3901 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3961 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4021 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4081 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4141 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4201 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4261 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4321 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4381 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4441 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4501 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4561 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4621 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5461 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5521 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5581 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5641 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     5701 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     5761 atgattacga attaattcga gctcgccc
//
