LOCUS       sequence-380728-        7905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GLUT8 protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7905)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 7905)
  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 GLUT8
FEATURES             Location/Qualifiers
     source          1..7905
                     /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(1719..1739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1724..2053
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1904..2039
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2101..2148
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2167..2565
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2652..2885
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2957..3078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2994..3013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3048..3067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3118..3253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3273..3292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3274..3292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3302..3319)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3302..3318)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3311..3333)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3460..3915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3547..3566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3757..3778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3941..4045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4046..4114,4115..4906)
                     /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(4264..4283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5077..5665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5566..5585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5819..5836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5953..5974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5989..5994,5995..6012,6013..6019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6027..6043
                     /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     6032..6054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6051..6067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6051..6067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6086..6106
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6088..6106
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6134..6360
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6361..6541
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6588..6713
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7206..7439
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7624..7668
                     /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 ctgcagatat ccaatgacgc
      841 ccgaggaccc gcgcttcctg ctgactcagc acaggcgcca ggaggccatg gccgccctgc
      901 ggttcctgtg gggctccgag cagggctggg aagacccccc catcggggct gagcagagct
      961 ttcacctggc cctgccgcgg cagcccggca tctacaagcc cttcatcatc ggcgtctccc
     1021 tgatggcctt ccagcagctg tcgggggtca acgccgtcat gttctatgca gagaccatct
     1081 ttgaagaggc caagttcaag gacagcagcc tggcctcggt cgtcgtgggt gtcatccagg
     1141 tgctgttcac agctgtggcg gctctcatca tggacagagc agggcggagg ctgctcctgg
     1201 tcttgtcagg tgtggtcatg gtgttcagca cgagtgcctt cggcgcctac ttcaagctga
     1261 cccagggtgg ccctggcaac tcctcgcacg tggccatctc ggcgcctgtc tctgcacagc
     1321 ctgttgatgc cagcgtgggg ctggcctggc tggccgtggg cagcatgtgc ctcttcatcg
     1381 ccggctttgc ggtgggctgg gggcccatcc cctggctcct catgtcagag atcttccctc
     1441 tgcatgtcaa gggcgtggcg acaggcatct gcgtcctcac caactggctc atggcctttc
     1501 tcgtgaccaa ggagttcagc agcctcatgg aggtcctcag gccctatgga gccttctggc
     1561 ttgcctccgc tttctgcatc ttcagtgtcc ttttcacttt gttctgtgtc cctgaaacta
     1621 aaggaaagac tctggaacaa atcacagccc attttgaggg gcgatgactc gagtctagag
     1681 ggcccaccgg ttagtaatga gtttggaatt aattctgtgg aatgtgtgtc agttagggtg
     1741 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1801 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1861 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1921 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1981 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2041 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     2101 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     2161 taaaccatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     2221 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     2281 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     2341 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     2401 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     2461 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     2521 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagcaca attcgagctc
     2581 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     2641 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     2701 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     2761 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     2821 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     2881 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     2941 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3001 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3061 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     3121 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3181 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3241 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     3301 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3361 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3421 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     3481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3901 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3961 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4021 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4081 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4141 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4201 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4261 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4321 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4381 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4441 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4501 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4561 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4621 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4681 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4741 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4801 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4921 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4981 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5041 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5641 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5701 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5761 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5821 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5881 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5941 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6001 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6061 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     6121 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     6181 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     6241 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     6301 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
     6361 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6421 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6481 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6541 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     6601 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     6661 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     6721 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     6781 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     6841 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     6901 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     6961 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     7021 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     7081 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     7141 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     7201 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     7261 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     7321 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     7381 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     7441 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     7501 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     7561 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     7621 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     7681 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     7741 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     7801 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     7861 gaagaagaag gtggagagag agacagagac agatccattc gatta
//
