LOCUS       sequence-177980-        7347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rglut1 HA complete.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7347)
  AUTHORS   Wieman HL, Wofford JA, Rathmell JC
  TITLE     Cytokine stimulation promotes glucose uptake via 
            phosphatidylinositol-3 kinase/Akt regulation of Glut1 activity and 
            trafficking.
  JOURNAL   Mol Biol Cell. 2007 Apr;18(4):1437-46. Epub 2007 Feb 14.
  PUBMED    17301289
REFERENCE   2  (bases 1 to 7347)
  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 Biol 
            Cell."; date: "2007-04"; volume: "18(4)"; pages: "1437-46. Epub 2007
            Feb 14"
COMMENT     Sequence Label: pEF6 rglut1 HA complete
FEATURES             Location/Qualifiers
     source          1..7347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1427..1453
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1460..1486
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2873..2914
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2924..2941
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2964..2981)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2970..3194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3240..3668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3327..3346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3537..3558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3677..3697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3682..4011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3862..3997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3924..3943
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4059..4106
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4125..4523
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4681..4802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4718..4737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4772..4791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4851..4867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4851..4867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4864..4886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4875..4891
                     /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(4899..4905,4906..4923,4924..4929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4944..4965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5082..5099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5322..5341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6001..6792,6793..6861))
                     /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     6624..6643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6862..6966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7041..7060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc gcagcgcggc
     1261 catggagccc agcagcaaga aggtgacggg ccgccttatg ttggccgtgg gaggggcagt
     1321 gctcggatcc ctgcagttcg gctataacac cggtgtcatc aacgcccccc agaaggtaat
     1381 tgaggagttc tacaatcaaa catggaacca ccgctatgga gagtcctatc cttatgacgt
     1441 tcctgactat gcagctagct atccttatga cgttcctgac tatgcaatcc catccaccac
     1501 actcaccaca ctctggtctc tctccgtggc catcttctct gtcgggggca tgattggttc
     1561 cttctctgtg ggcctctttg ttaatcgctt tggcaggcgg aactccatgc tgatgatgaa
     1621 cctgttggcc tttgtgtctg ccgtgcttat gggtttctcc aaactgggca agtcctttga
     1681 gatgctgatc ctgggccgct tcatcattgg agtgtactgt ggcctgacca ccggctttgt
     1741 gcccatgtat gtgggggagg tgtcacccac agctcttcgt ggagccctgg gcaccctgca
     1801 ccagctgggc atcgtcgttg ggatccttat tgcccaggtg ttcggcttag actccatcat
     1861 gggcaatgca gacttgtggc ctctactgct cagtgtcatc ttcatcccag ccctgctaca
     1921 gtgtatcctg ttgcccttct gccctgagag cccccgcttc ctgctcatca atcgtaacga
     1981 ggagaaccgg gccaagagtg tgctgaaaaa gcttcgaggg acagccgatg tgacccgaga
     2041 cctgcaggag atgaaagaag agggtcggca gatgatgcgg gagaagaagg tcaccatctt
     2101 ggagctgttc cgctcacccg cctaccgcca gcccatcctc atcgccgtgg tgctgcagct
     2161 gtcccagcag ctgtcgggca tcaatgctgt gttctactac tcaacgagca tcttcgagaa
     2221 ggcaggtgtg cagcagcctg tgtatgccac catcggctcg ggtatcgtca acacggcctt
     2281 cactgtggtg tcgctgttcg tcgtggagcg agctggccgt cggaccctgc acctcattgg
     2341 tctggctggc atggcgggct gtgctgtgct catgaccatc gccctggccc tgctggagca
     2401 gctgccctgg atgtcctatc tgagtatcgt ggccatcttt ggctttgtgg ccttctttga
     2461 agtaggccct ggtcctattc catggttcat tgtggccgag ctgttcagcc aggggccccg
     2521 acctgctgct gttgctgtgg ctggcttctc taactggacc tcaaacttca tcgtgggcat
     2581 gtgcttccaa tatgtggagc aactgtgtgg cccctacgtc ttcatcatct tcacggtgct
     2641 gctggtactc ttcttcatct tcacctactt caaagttcct gagaccaaag gccggacctt
     2701 cgatgagatc gcttccggct tccggcaggg gggtgccagc cagagcgaca agacacctga
     2761 ggagctcttc caccctctgg gggctgactc ccaagtgtga ggagccaagg gcaattctgc
     2821 agatatccag cacagtggcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc
     2881 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     2941 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3001 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3061 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3121 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3181 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3241 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3301 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3361 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3421 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3481 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3541 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3601 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3661 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4021 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     4081 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagcctttgt
     4141 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     4201 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     4261 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     4321 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     4381 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     4441 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     4501 ctggttatgt gtgggagggc taagcacttc gtggccgagg agcaggactg acacgtgcta
     4561 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4621 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4681 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4741 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4801 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4861 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4921 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4981 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5041 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5161 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7021 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7081 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7141 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     7201 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag gcttttgcaa
     7261 aaagctttgc aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc
     7321 ttctaggtct tgaaaggagt gggaatt
//
