LOCUS       sequence-160459-        7219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  detection and measurement of GLUT4 translocation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7219)
  AUTHORS   Bogan JS, McKee AE, Lodish HF
  TITLE     Insulin-responsive compartments containing GLUT4 in 3T3-L1 and CHO 
            cells: regulation by amino acid concentrations.
  JOURNAL   Mol Cell Biol. 2001 Jul;21(14):4785-806.
  PUBMED    11416153
REFERENCE   2  (bases 1 to 7219)
  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: "Mol Cell 
            Biol."; date: "2001-07"; volume: "21(14)"; pages: "4785-806"
COMMENT     Sequence Label: pB-GLUT4-7myc-GFP
FEATURES             Location/Qualifiers
     source          1..7219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..594
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    657..1014
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1079..1495
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1425..1447
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1463..1479
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1505..1879
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1817..1833
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             2126..2155
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2165..2194
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2204..2233
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2243..2272
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2282..2311
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2345..2374
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2402..2431
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(3582..3593)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3788..3797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3794..3796,3797..3799,3800..4513)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3794..3796,3797..3799,3800..4510)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3839..3860)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4100..4119)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4447..4468
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4703..5172
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5302..5319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5473..6061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5553..5572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6225..7016,7017..7085))
                     /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     6848..6867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7086..7190)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
      121 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      181 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      241 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      301 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      361 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      421 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      481 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      541 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      601 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      661 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      721 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      781 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      841 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt
      901 agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      961 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1021 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt tgggcccggg
     1081 ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat
     1141 cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga
     1201 atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac
     1261 ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta
     1321 catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca
     1381 ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg
     1441 ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat
     1501 atggccatat gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga
     1561 catgacaaga gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt
     1621 ccagcacgaa gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt
     1681 ggtacctcac ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa
     1741 cctagaacct cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa
     1801 agtagacggc atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg
     1861 tggaccatcc tctagactgc cggatcccag tgtggtggta cgggaattct aagccgccac
     1921 catgccgtcg ggtttccagc agatcggctc tgaagatggg gaaccccctc agcagcgagt
     1981 gactgggaca ctggtccttg ctgtattctc agctgtgctt ggctcccttc agtttggcta
     2041 taacattgga gtcatcaacg ccccacagaa agtgattgaa cagagctaca atgcaacttg
     2101 gctgggtagg cagggtcctg cagaggagca aaagcttatt tctgaagagg acttgaatga
     2161 aatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc
     2221 tgaagaggac ttgaatgaaa tggagcaaaa gctcatttct gaagaggact tgaatgaaat
     2281 ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagagcttgg gcgacctcac
     2341 catggagcaa aagctcattt ctgaagagga cttgaactca agacctcttg agcctgcaga
     2401 ggagcaaaag cttatttctg aagaggactt gcttaaggga cccagctcca tccctccagg
     2461 caccctcacc accctctggg ccctctccgt ggccatcttt tccgtgggcg gcatgatttc
     2521 ctccttcctc attggtatca tctctcagtg gcttggaagg aaaagggcca tgctggtcaa
     2581 caatgtcctg gcggtgctgg ggggcagcct catgggcctg gccaacgctg ctgcctccta
     2641 tgaaatgctc atccttggac gattcctcat tggcgcctac tcagggctga catcagggct
     2701 ggtgcccatg tacgtggggg agattgctcc cactcacctg cggggcgccc tggggacgct
     2761 caaccaactg gccattgtta tcggcattct gatcgcccag gtgctgggct tggagtccct
     2821 cctgggcact gccagcctgt ggccactgct cctgggcctc acagtgctac ctgccctcct
     2881 gcagctggtc ctgctgccct tctgtcccga gagcccccgc tacctctaca tcatccagaa
     2941 tctcgagggg cctgccagaa agagtctgaa gcgcctgaca ggctgggccg atgtttctgg
     3001 agtgctggct gagctgaagg atgagaagcg gaagctggag cgtgagcggc cactgtccct
     3061 gctccagctc ctgggcagcc gtacccaccg gcagcccctg atcattgcgg tcgtgctgca
     3121 gctgagccag cagctctctg gcatcaatgc tgttttctat tattcgacca gcatcttcga
     3181 gacagcaggg gtaggccagc ctgcctatgc caccatagga gctggtgtgg tcaacacagt
     3241 cttcaccttg gtctcggtgt tgttggtgga gcgggcgggg cgccggacgc tccatctcct
     3301 gggcctggcg ggcatgtgtg gctgtgccat cctgatgact gtggctctgc tcctgctgga
     3361 gcgagttcca gccatgagct acgtctccat tgtggccatc tttggcttcg tggcattttt
     3421 tgagattggc cctggcccca ttccttggtt catcgtggcc gagctcttca gccagggacc
     3481 ccgcccggca gccatggctg tggctggttt ctccaactgg acgagcaact tcatcattgg
     3541 catgggtttc cagtatgttg cggaggctat ggggccctac gtcttccttc tatttgcggt
     3601 cctcctgctg ggcttcttca tcttcacctt cttaagagta cctgaaactc gaggccggac
     3661 gtttgaccag atctcggctg ccttccaccg gacaccctct cttttagagc aggaggtgaa
     3721 acccagcaca gaacttgagt atttagggcc agatgagaat gacccgcggg cccgggatcc
     3781 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3841 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3901 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3961 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     4021 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4081 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     4141 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4201 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     4261 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     4321 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4381 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     4441 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     4501 gctgtacaag taaagcggcc gccagcacag tggtcgacga taaaataaaa gattttattt
     4561 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4621 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca
     4681 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     4741 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
     4801 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4861 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     4921 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     4981 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct
     5041 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     5101 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     5161 gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     5221 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5281 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5341 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5401 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5461 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5521 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5581 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5641 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5701 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5761 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5821 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5881 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5941 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6001 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6061 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6121 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc
     6181 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7081 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7141 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7201 cgaaaagtgc cacctgcat
//
