LOCUS       sequence-380736-        8583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GLUT6 protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8583)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8583)
  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"
COMMENT     Sequence Label: pLenti6MQBI163 GLUT6
FEATURES             Location/Qualifiers
     source          1..8583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        346..549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     500..520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     546..570
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    626..788
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             792..815
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2397..2417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2402..2731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2582..2717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2779..2826
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2845..3243
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3330..3563
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3635..3756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3672..3691)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3726..3745
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3796..3931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3951..3970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3952..3970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3980..3997)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3980..3996)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3989..4011)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4138..4593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4225..4244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4435..4456
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4619..4723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4724..4792,4793..5584)
                     /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(4942..4961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5755..6343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6244..6263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6497..6514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6631..6652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6667..6672,6673..6690,6691..6697)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6705..6721
                     /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     6710..6732
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6729..6745
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6729..6745
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6764..6784
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6766..6784
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6812..7038
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7039..7219
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7266..7391
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7884..8117
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8302..8346
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 gtgaacggat ctcgacggta tcgataagct tgggagttcc gcgttacata acttacggta
       61 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      121 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      181 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      241 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      301 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      361 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      421 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      481 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      541 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
      601 ctccatagaa gacaccgact ctagtagcgc agaggcttgg ggcagccgag cggcagccag
      661 gccccggccc gggcctcggt tccagaaggg agaggagccc gccaaggcgc gcaagagagc
      721 gggctgcctc gcagtccgag ccggagaggg agcgcgagcc gcgccggccc cggacggcct
      781 ccgaaaccat ggattacaaa gacgatgacg acaaggaatt catgcaggag ccgctgctgg
      841 gagccgaggg cccggactac gacaccttcc ccgagaagcc gcccccgtcg ccaggggaca
      901 gggcgcgggt cgggaccctg cagaacaaaa gggtgttcct ggccaccttc gccgcagtgc
      961 tcggcaattt cagctttggg tatgccctgg tctacacatc ccctgtcatc ccagccctgg
     1021 agcgctcctt ggatcctgac ctgcatctga ccaaatccca ggcatcctgg tttgggtccg
     1081 tgttcaccct gggagcagcg gccggaggcc tgagtgccat gatcctcaac gacctcctgg
     1141 gccggaagct gagcatcatg ttctcagctg tgccgtcggc ggccggctat gcgctcatgg
     1201 cgggtgcgca cggcctctgg atgctgctgc tcggaaggac gctgacgggc ttcgccgggg
     1261 ggctcacagc tgcctgcatc ccggtgtacg tgtctgagat tgctccccca ggcgttcgtg
     1321 gggctctggg ggccacaccc cagctcatgg cagtgttcgg atccctgtcc ctctacgccc
     1381 ttggcctcct gctgccgtgg cgctggctgg ctgtggccgg ggaggcgcct gtgctcatca
     1441 tgatcctgct gctcagcttc atgcccaact cgccgcgctt cctgctctct cggggcaggg
     1501 acgaagaggc cctgcgggcg ctggcctggc tgcgtgggac ggacgtcgat gtccactggg
     1561 agttcgagca gatccaggac aacgtccgga gacagagcag ccgagtatcg tgggctgagg
     1621 cacgggcccc ccacgtgtgc cggcccatca ccgtggcctt gctgatgcgc ctcctgcagc
     1681 agctgacggg catcacgccc atcctggtct acctgcagtc catcttcgac agcaccgctg
     1741 tcctgctgcc ccccaaggac gacgcagcca tcgttggggc cgtgcggctc ctgtccgtgc
     1801 tgatcgccgc cctcaccatg gacctcgcag gccgcaaggt gctgctcttc gtctcagcgg
     1861 ccatcatgtt tgctgccaac ctgactctgg ggctgtacat ccactttggc cccaggcctc
     1921 tgagccccaa cagcactgcg ggcctggaaa gcgagtcctg gggggacttg gcgcagcccc
     1981 tggcagcacc cgctggctac ctcaccctgg tgcccctgct ggccaccatg ctcttcatca
     2041 tgggctacgc cgtgggctgg ggtcccatca cctggctgct catgtctgag gtcctgcccc
     2101 tgcgtgcccg tggcgtggcc tcagggctct gcgtgctggc cagctggctc accgccttcg
     2161 tcctcaccaa gtccttcctg ccagtggtga gcaccttcgg cctccaggtg cctttcttct
     2221 tcttcgcggc catctgcttg gtgagcctgg tgttcacagg ctgctgtgtg cccgagacca
     2281 agggacggtc cctggagcag atcgagtcct tcttccgcac ggggagaagg tccttcttgc
     2341 gctagctcga gtctagaggg cccaccggtt agtaatgagt ttggaattaa ttctgtggaa
     2401 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2461 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2521 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2641 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2701 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     2761 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     2821 tacgacaagg tgaggaacta aaccatggcc aagcctttgt ctcaagaaga atccaccctc
     2881 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     2941 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     3001 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     3061 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     3121 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     3181 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     3241 taagcacaat tcgagctcgg tacctttaag accaatgact tacaaggcag ctgtagatct
     3301 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     3361 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     3421 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     3481 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     3541 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     3601 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     3661 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3721 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     3781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3901 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     3961 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4021 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4081 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     4141 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4201 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4261 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4321 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4381 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4441 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4501 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4561 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5881 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6421 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6481 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6541 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6601 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6661 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6721 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     6781 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     6841 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     6901 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
     6961 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
     7021 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7081 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7141 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7201 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     7261 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     7321 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     7381 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     7441 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     7501 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     7561 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     7621 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     7681 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     7741 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     7801 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     7861 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     7921 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     7981 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     8041 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     8101 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     8161 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     8221 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     8281 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     8341 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     8401 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     8461 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     8521 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     8581 tta
//
