LOCUS       sequence-175400-        8106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GLUT1 construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8106)
  AUTHORS   Wieman HL, Horn SR, Jacobs SR, Altman BJ, Kornbluth S, Rathmell JC
  TITLE     An essential role for the Glut1 PDZ-binding motif in growth factor 
            regulation of Glut1 degradation and trafficking.
  JOURNAL   Biochem J. 2009 Mar 1;418(2):345-67. doi: 10.1042/BJ20081422.
  PUBMED    19016655
REFERENCE   2  (bases 1 to 8106)
  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"; journalName: 
            "Biochem J"; date: "2009-03-1- 1"; volume: "418"; issue: "2"; pages:
            "345-67"
COMMENT     Sequence Label: GLUT1-GFP
FEATURES             Location/Qualifiers
     source          1..8106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(34..62)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     657..677
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2338..2379
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2389..2406
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2429..2446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2435..2659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2705..3133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2792..2811)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3002..3023
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3142..3162)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3147..3476
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3327..3462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3389..3408
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3524..3571
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3590..3988
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4146..4267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4183..4202)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4237..4256
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4316..4332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4316..4332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4329..4351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4340..4356
                     /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(4364..4370,4371..4388,4389..4394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4409..4430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4547..4564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4707..5295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4787..4806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5466..6257,6258..6326))
                     /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     6089..6108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6327..6431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6506..6525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6813..7991
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7044..7982
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7939..7959
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     8008..8027
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8008..8026
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atggccagca aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
       61 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgctacatac
      121 ggaaagctta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
      181 cttgtcacta ctttctctta tggtgttcaa tgcttttccc gttatccgga tcatatgaaa
      241 cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct
      301 ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg tgataccctt
      361 gttaatcgta tcgagttaaa aggtattgat tttaaagaag atggaaacat tctcggacac
      421 aaactcgagt acaactataa ctcacacaat gtatacatca cggcagacaa acaaaagaat
      481 ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggatccgt tcaactagca
      541 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
      601 tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc
      661 cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaaagc
      721 ggttccggac cggtgctagc ggtaccgagc tcggatccac tagtccagtg tggtggaatt
      781 gcccttatgg agcccagcag caagaaggtg acgggccgcc ttatgttggc cgtgggaggg
      841 gcagtgctcg gatccctgca gttcggctat aacaccggtg tcatcaacgc cccccagaag
      901 gtaattgagg agttctacaa tcaaacatgg aaccaccgct atggagagtc catcccatcc
      961 accacactca ccacactctg gtctctctcc gtggccatct tctctgtcgg gggcatgatt
     1021 ggttccttct ctgtgggcct ctttgttaat cgctttggca ggcggaactc catgctgatg
     1081 atgaacctgt tggcctttgt gtctgccgtg cttatgggtt tctccaaact gggcaagtcc
     1141 tttgagatgc tgatcctggg ccgcttcatc attggagtgt actgtggcct gaccaccggc
     1201 tttgtgccca tgtatgtggg ggaggtgtca cccacagctc ttcgtggagc cctgggcacc
     1261 ctgcaccagc tgggcatcgt cgttgggatc cttattgccc aggtgttcgg cttagactcc
     1321 atcatgggca atgcagactt gtggcctcta ctgctcagtg tcatcttcat cccagccctg
     1381 ctacagtgta tcctgttgcc cttctgccct gagagccccc gcttcctgct catcaatcgt
     1441 aacgaggaga accgggccaa gagtgtgctg aaaaagcttc gagggacagc cgatgtgacc
     1501 cgagacctgc aggagatgaa agaagagggt cggcagatga tgcgggagaa gaaggtcacc
     1561 atcttggagc tgttccgctc acccgcctac cgccagccca tcctcatcgc cgtggtgctg
     1621 cagctgtccc agcagctgtc gggcatcaat gctgtgttct actactcaac gagcatcttc
     1681 gagaaggcag gtgtgcagca gcctgtgtat gccaccatcg gctcgggtat cgtcaacacg
     1741 gccttcactg tggtgtcgct gttcgtcgtg gagcgagctg gccgtcggac cctgcacctc
     1801 attggtctgg ctggcatggc gggctgtgct gtgctcatga ccatcgccct ggccctgctg
     1861 gagcagctgc cctggatgtc ctatctgagt atcgtggcca tctttggctt tgtggccttc
     1921 tttgaagtag gccctggtcc tattccatgg ttcattgtgg ccgagctgtt cagccagggg
     1981 ccccgacctg ctgctgttgc tgtggctggc ttctctaact ggacctcaaa cttcatcgtg
     2041 ggcatgtgct tccaatatgt ggagcaactg tgtggcccct acgtcttcat catcttcacg
     2101 gtgctgctgg tactcttctt catcttcacc tacttcaaag ttcctgagac caaaggccgg
     2161 accttcgatg agatcgcttc cggcttccgg caggggggtg ccagccagag cgacaagaca
     2221 cctgaggagc tcttccaccc tctgggggct gactcccaag tgtgaggagc caagggcaat
     2281 tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgcg gttcgaaggt
     2341 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat
     2401 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2461 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2521 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2581 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2641 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2701 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2761 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2821 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2881 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2941 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3001 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3061 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3121 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3241 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3301 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3361 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3421 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3481 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     3541 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagcc
     3601 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     3661 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     3721 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     3781 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     3841 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     3901 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     3961 gccctctggt tatgtgtggg agggctaagc acttcgtggc cgaggagcag gactgacacg
     4021 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4081 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4141 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4201 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4261 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccaggaac
     4681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6421 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     6481 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     6541 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     6601 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     6661 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actaggcttt
     6721 tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat
     6781 ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     6841 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     6901 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     6961 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     7021 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
     7081 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc
     7141 ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag
     7201 ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa
     7261 tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt
     7321 tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag
     7381 atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc
     7441 agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt
     7501 agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc
     7561 cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc
     7621 ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg
     7681 agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca
     7741 cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt
     7801 ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga
     7861 ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt
     7921 ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc
     7981 aggtgtcgtg aggaattagc ttggtactaa tacgactcac tatagggaga cccaagctgg
     8041 ctaggtaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa ttgcccttta
     8101 gacacc
//
