LOCUS       sequence-315359-        6271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal HA-tagged GLUT5 cDNA expression vector for transfections.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6271)
  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 6271)
  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: HA-GLUT5 in pRK5
FEATURES             Location/Qualifiers
     source          1..6271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    82..103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(118..123,124..141,142..148)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    156..172
                     /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     161..183
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     180..196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     180..196
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        225..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        605..808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     805..829
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1039..1057
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1039..1056
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1070..1089
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1174..1183
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1186..1212
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2782..2791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2795..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2886..2905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2998..3355
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2998..3189
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3021..3041)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3206..3341
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3375..3392)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3375..3391)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3384..3406)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3604..4059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3691..3710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3901..3922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4076..4095)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4195..4217
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4255..4273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4341..4445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4446..4514,4515..5306)
                     /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(4664..4683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5477..6065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5966..5985
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6219..6236
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
       61 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      121 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      181 aggaaacagc tatgacatga ttacgaatta attcgagctc gcccgacatt gattattgac
      241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      841 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
      901 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
      961 agtctatagg cccaccccct tggcttcgtt agaacgcggc tacaattaat acataacctt
     1021 atgtatcata cacatacgat ttaggtgaca ctatagaata acatccactt tgcctttctc
     1081 tccacaggtg tccactccca ggtccaactg cacctcggtt ctatcgattg aattcccgaa
     1141 ccgacagtcg gtctcttcac caaggccatt cgcgccacca tgggctaccc ctatgatgtg
     1201 cctgactacg cagatctcaa tggtggtggt ggtgggtcga cgatggagca acaggatcag
     1261 agcatgaagg aagggaggct gacgcttgtg cttgccctgg caaccctgat agctgccttt
     1321 gggtcatcct tccagtatgg gtacaacgtg gctgctgtca actccccagc actgctcatg
     1381 caacaatttt acaatgagac ttactatggt aggaccggtg aattcatgga agacttcccc
     1441 ttgacgttgc tgtggtctgt aaccgtgtcc atgtttccat ttggagggtt tatcggatcc
     1501 ctcctggtcg gccccttggt gaataaattt ggcagaaaag gggccttgct gttcaacaac
     1561 atattttcta tcgtgcctgc gatcttaatg ggatgcagca gagtcgccac atcatttgag
     1621 cttatcatta tttccagact tttggtggga atatgtgcag gtgtatcttc caacgtggtc
     1681 cccatgtact taggggagct ggcccctaaa aacctgcggg gggctctcgg ggtggtgccc
     1741 cagctcttca tcactgttgg catccttgtg gcccagatct ttggtcttcg gaatctcctt
     1801 gcaaacgtag atggctggcc gatcctgctg gggctgaccg gggtccccgc ggcgctgcag
     1861 ctccttctgc tgcccttctt ccccgagagc cccaggtacc tgctgattca gaagaaagac
     1921 gaagcggccg ccaagaaagc cctacagacg ctgcgcggct gggactctgt ggacagggag
     1981 gtggccgaga tccggcagga ggatgaggca gagaaggccg cgggcttcat ctccgtgctg
     2041 aagctgttcc ggatgcgctc gctgcgctgg cagctgctgt ccatcatcgt cctcatgggc
     2101 ggccagcagc tgtcgggcgt caacgctatc tactactacg cggaccagat ctacctgagc
     2161 gccggcgtgc cggaggagca cgtgcagtac gtgacggccg gcaccggggc cgtgaacgtg
     2221 gtcatgacct tctgcgccgt gttcgtggtg gagctcctgg gtcggaggct gctgctgctg
     2281 ctgggcttct ccatctgcct catagcctgc tgcgtgctca ctgcagctct ggcactgcag
     2341 gacacagtgt cctggatgcc atacatcagc atcgtctgtg tcatctccta cgtcatagga
     2401 catgccctcg ggcccagtcc catacccgcg ctgctcatca ctgagatctt cctgcagtcc
     2461 tctcggccat ctgccttcat ggtggggggc agtgtgcact ggctctccaa cttcaccgtg
     2521 ggcttgatct tcccgttcat ccaggagggc ctcggcccgt acagcttcat tgtcttcgcc
     2581 gtgatctgcc tcctcaccac catctacatc ttcttgattg tcccggagac caaggccaag
     2641 acgttcatag agatcaacca gattttcacc aagatgaata aggtgtctga agtgtacccg
     2701 gaaaaggagg aactgaaaga gcttccacct gtcacttcgg aacagtgagc ggccgctaag
     2761 taagtaagga tccccagctt ggccgccatg gcccaacttg tttattgcag cttataatgg
     2821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2881 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg ggaattaatt
     2941 cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3001 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3061 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3181 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3241 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3301 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3361 gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3421 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3481 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3541 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     3601 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3661 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3721 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3781 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     3841 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3901 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     3961 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4021 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4081 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4141 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4201 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4261 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4321 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4381 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4441 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4501 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4561 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4621 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4681 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4741 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4801 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4861 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4921 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4981 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5041 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5101 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5161 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5221 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5281 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5341 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5401 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5461 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5521 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5581 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5641 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5701 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5761 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5821 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5881 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5941 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6001 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6061 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6121 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6181 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6241 cggaagagcg cccaatacgc aaaccgcctc t
//
