LOCUS       sequence-380702-        8127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GLUT2 protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8127)
  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 GLUT2
FEATURES             Location/Qualifiers
     source          1..8127
                     /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(1941..1961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1946..2275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2126..2261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2323..2370
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2389..2787
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2874..3107
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3179..3300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3216..3235)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3270..3289
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3340..3475
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3495..3514)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3496..3514)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3524..3541)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3524..3540)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3533..3555)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3682..4137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3769..3788)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3979..4000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4163..4267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4268..4336,4337..5128)
                     /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(4486..4505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5299..5887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5788..5807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6041..6058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6175..6196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6211..6216,6217..6234,6235..6241)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6249..6265
                     /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     6254..6276
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6273..6289
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6273..6289
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6308..6328
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6310..6328
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6356..6582
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6583..6763
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6810..6935
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7428..7661
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7846..7890
                     /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 ccaatgtata
      841 tcggtgaaat tgctccaacc gctctcaggg gagcacttgg cacttttcat cagctggcca
      901 tcgtcacggg cattcttatt agtcagatta ttggtcttga atttatcttg ggcaattatg
      961 atctgtggca catcctgctt ggcctgtctg gtgtgcgagc catccttcag tctctgctac
     1021 tctttttctg tccagaaagc cccagatacc tttacatcaa gttagatgag gaagtcaaag
     1081 caaaacaaag cttgaaaaga ctcagaggat atgatgatgt caccaaagat attaatgaaa
     1141 tgagaaaaga aagagaagaa gcatcgagtg agcagaaagt ctctataatt cagctcttca
     1201 ccaattccag ctaccgacag cctattctag tggcactgat gctgcatgtg gctcagcaat
     1261 tttccggaat caatggcatt ttttactact caaccagcat ttttcagacg gctggtatca
     1321 gcaaacctgt ttatgcaacc attggagttg gcgctgtaaa catggttttc actgctgtct
     1381 ctgtattcct tgtggagaag gcagggcgac gttctctctt tctaattgga atgagtggga
     1441 tgtttgtttg tgccatcttc atgtcagtgg gacttgtgct gctgaataag ttctcttgga
     1501 tgagttatgt gagcatgata gccatcttcc tctttgtcag cttctttgaa attgggccag
     1561 gcccgatccc ctggttcatg gtggctgagt ttttcagtca aggaccacgt cctgctgctt
     1621 tagcaatagc tgcattcagc aattggacct gcaatttcat tgtagctctg tgtttccagt
     1681 acattgcgga cttctgtgga ccttatgtgt ttttcctctt tgctggagtg ctcctggcct
     1741 ttaccctgtt cacatttttt aaagttccag aaaccaaagg aaagtctttt gaggaaattg
     1801 ctgcagaatt ccaaaagaag agtggctcag cccacaggcc aaaagctgct gtagaaatga
     1861 aattcctagg agctacagag actgtgtaac tcgagtctag agggcccacc ggttagtaat
     1921 gagtttggaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1981 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2041 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2101 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2161 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2221 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2281 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     2341 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct
     2401 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     2461 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     2521 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     2581 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     2641 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     2701 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     2761 ccctctggtt atgtgtggga gggctaagca caattcgagc tcggtacctt taagaccaat
     2821 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     2881 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     2941 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     3001 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     3061 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     3121 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     3181 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3241 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3301 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
     3361 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3421 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
     3481 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     3541 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3601 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3661 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3721 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3781 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3841 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3901 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3961 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4021 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4081 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     4141 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4201 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4261 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4321 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4381 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4441 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4501 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4561 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4621 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4681 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4741 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4801 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4861 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4921 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4981 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5041 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5101 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5161 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5221 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5281 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5341 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5401 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5461 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5521 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5581 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5641 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5701 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5761 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5821 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5881 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5941 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6001 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6061 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6121 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6181 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6241 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     6301 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag
     6361 tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta
     6421 caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc
     6481 ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat
     6541 tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac
     6601 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6661 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6721 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     6781 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     6841 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     6901 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
     6961 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     7021 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     7081 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     7141 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     7201 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     7261 ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     7321 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     7381 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     7441 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     7501 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     7561 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     7621 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     7681 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     7741 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     7801 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     7861 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     7921 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     7981 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     8041 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     8101 agagacagag acagatccat tcgatta
//
