LOCUS       sequence-401489-        8713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT human GLUT1 with a C-terminal 3xFLAG tag in pQCXIP vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8713)
  AUTHORS   Qualls-Histed SJ, Nielsen CP, MacGurn JA
  TITLE     Lysosomal trafficking of the glucose transporter GLUT1 requires 
            sequential regulation by TXNIP and ubiquitin.
  JOURNAL   iScience. 2023 Feb 6;26(3):106150. doi: 10.1016/j.isci.2023.106150. 
            eCollection 2023 Mar 17.
  PUBMED    36890792
REFERENCE   2  (bases 1 to 8713)
  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"; journalName: "iScience.";
            date: "2023-02-6"; volume: "26(3):106150. doi"; pages: " 
            10.1016/j.isci.2023.106150. eCollection 2023 Mar 17"
COMMENT     Sequence Label: WT GLUT1-FLAG
FEATURES             Location/Qualifiers
     source          1..8713
                     /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"
     LTR             585..760
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    823..1022
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1223..1639
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1569..1591
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1607..1623
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1676..1979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1980..2183
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2180..2204
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3819..3884
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3912..4486,4487..4489,4490..4498)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4079..4096)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4306..4325
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4518..5117
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4518..4537)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5014..5034
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5491..5916
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6019..6041)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6177..6197)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6182..6511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6362..6497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6533..6552
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6633..6650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6804..7392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6884..6903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7563..8354,8355..8423))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8186..8205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8424..8528)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8596..8614
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 agctcaataa aagagcccac
      601 aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc
      661 caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct
      721 cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt
      781 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
      841 tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta
      901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      961 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
     1021 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     1081 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     1141 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1201 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1261 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1321 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1381 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1441 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1501 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1561 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1621 cactccttct ctaggcgccg gaattgaaga tctgggggat cgatcctcta gagtccgtta
     1681 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1741 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1801 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1861 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1921 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1981 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     2041 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     2101 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     2161 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     2221 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgggaac
     2281 ggtgcattgg aacggacctg caggaattga tccgcggccg catggagccc agcagcaaga
     2341 agctgacggg tcgcctcatg ctggccgtgg gaggagcagt gcttggctcc ctgcagtttg
     2401 gctacaacac tggagtcatc aatgcccccc agaaggtgat cgaggagttc tacaaccaga
     2461 catgggtcca ccgctatggg gagagcatcc tgcccaccac gctcaccacg ctctggtccc
     2521 tctcagtggc catcttttct gttgggggca tgattggctc cttctctgtg ggccttttcg
     2581 ttaaccgctt tggccggcgg aattcaatgc tgatgatgaa cctgctggcc ttcgtgtccg
     2641 ccgtgctcat gggcttctcg aaactgggca agtcctttga gatgctgatc ctgggccgct
     2701 tcatcatcgg tgtgtactgc ggcctgacca caggcttcgt gcccatgtat gtgggtgaag
     2761 tgtcacccac agcccttcgt ggggccctgg gcaccctgca ccagctgggc atcgtcgtcg
     2821 gcatcctcat cgcccaggtg ttcggcctgg actccatcat gggcaacaag gacctgtggc
     2881 ccctgctgct gagcatcatc ttcatcccgg ccctgctgca gtgcatcgtg ctgcccttct
     2941 gccccgagag tccccgcttc ctgctcatca accgcaacga ggagaaccgg gccaagagtg
     3001 tgctaaagaa gctgcgcggg acagctgacg tgacccatga cctgcaggag atgaaggaag
     3061 agagtcggca gatgatgcgg gagaagaagg tcaccatcct ggagctgttc cgctcccccg
     3121 cctaccgcca gcccatcctc atcgctgtgg tgctgcagct gtcccagcag ctgtctggca
     3181 tcaacgctgt cttctattac tccacgagca tcttcgagaa ggcgggggtg cagcagcctg
     3241 tgtatgccac cattggctcc ggtatcgtca acacggcctt cactgtcgtg tcgctgtttg
     3301 tggtggagcg agcaggccgg cggaccctgc acctcatagg cctcgctggc atggcgggtt
     3361 gtgccatact catgaccatc gcgctagcac tgctggagca gctaccctgg atgtcctatc
     3421 tgagcatcgt ggccatcttt ggctttgtgg ccttctttga agtgggtcct ggccccatcc
     3481 catggttcat cgtggctgaa ctcttcagcc agggtccacg tccagctgcc attgccgttg
     3541 caggcttctc caactggacc tcaaatttca ttgtgggcat gtgcttccag tatgtggagc
     3601 aactgtgtgg tccctacgtc ttcatcatct tcactgtgct cctggttctg ttcttcatct
     3661 tcacctactt caaagttcct gagactaaag gccggacctt cgatgagatc gcttccggct
     3721 tccggcaggg gggagccagc caaagtgaca agacacccga ggagctgttc catcccctgg
     3781 gggctgattc ccaagtggga tcctcaagtt tgccgggcga ctataaagac cacgacggtg
     3841 actacaagga ccacgatata gattacaaag atgacgacga taaatgagga attccgcccc
     3901 cccccccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     3961 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     4021 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     4081 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     4141 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     4201 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     4261 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     4321 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     4381 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     4441 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaaccca
     4501 caaggagacg accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     4561 acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     4621 acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     4681 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     4741 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     4801 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     4861 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     4921 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     4981 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     5041 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     5101 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     5161 gaccccatgg ctccgaccga agccacccgg ggcggccccg ccgaccccgc acccgccccc
     5221 gaggcccacc gactctagtc gagggctgca gcgctgcaga ggccgagtgc agaactgctc
     5281 caaagggacc tcaaggcttt ccgagggaca ctaggctgac tccatcgagc cagtgtagag
     5341 ataagcttat cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     5401 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     5461 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     5521 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     5581 gttcagatca aggtcaggaa cagatggaac agggtcgacc ctagagaacc atcagatgtt
     5641 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     5701 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     5761 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     5821 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     5881 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag
     5941 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt
     6001 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6061 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6121 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtagatcc ggctgtggaa
     6181 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6241 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     6301 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6361 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6421 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6481 aggctttttt ggaggcctag gcttttgcaa aaagcttact ggcttaacta tgcggcatca
     6541 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     6601 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7201 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7861 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8161 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     8221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8521 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8581 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attagcttgg
     8641 ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca
     8701 ttaccgccat gtt
//
