LOCUS       sequence-380731-        8598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GLUT9 protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8598)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8598)
  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: pLenti6M GLUT9
FEATURES             Location/Qualifiers
     source          1..8598
                     /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"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             641..664
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             683..703
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             707..748
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2334..2375
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2412..2432)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2417..2746
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2597..2732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2794..2841
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2860..3258
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3345..3578
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3650..3771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3687..3706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3741..3760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3811..3946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3966..3985)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3967..3985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3995..4012)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3995..4011)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4004..4026)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4153..4608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4240..4259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4450..4471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4634..4738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4739..4807,4808..5599)
                     /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(4957..4976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5770..6358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6259..6278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6512..6529
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6646..6667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6682..6687,6688..6705,6706..6712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6720..6736
                     /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     6725..6747
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6744..6760
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6744..6760
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6779..6799
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6781..6799
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6827..7053
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7054..7234
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7281..7406
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7899..8132
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8317..8361
                     /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 ctagaggatc cgccaccatg gattacaagg atgacgacga
      661 taagagcccg ggcggatcgt tggagaatct ttattttcag ggtcaaggta agcctatccc
      721 taaccctctc ctcggtctcg attctacgtc tagactgcag gaattcgata tcatgaagct
      781 cagtaaaaag gaccgaggag aagatgaaga aagtgattca gcgaaaaaga aattggactg
      841 gtcctgctcg ctcctcgtgg cctccctcgc gggcgccttc ggctcctcct tcctctacgg
      901 ctacaacctg tcggtggtga atgcccccac cccgtacatc aaggcctttt acaatgagtc
      961 atgggaaaga aggcatggac gtccaataga cccagacact ctgactctgc tctggtctgt
     1021 gactgtgtcc atattcgcca tcggtggact tgtggggaca ttaattgtga agatgattgg
     1081 aaaggttctt gggaggaagc acactttgct ggccaataat gggtttgcaa tttctgctgc
     1141 attgctgatg gcctgctcgc tccaggcagg agcctttgaa atgctcatcg tgggacgctt
     1201 catcatgggc atagatggag gcgtcgccct cagtgtgctc cccatgtacc tcagtgagat
     1261 ctcacccaag gagatccgtg gctctctggg gcaggtgact gccatcttta tctgcattgg
     1321 cgtgttcact gggcagcttc tgggcctgcc cgagctgctg ggaaaggaga gtacctggcc
     1381 atacctgttt ggagtgattg tggtccctgc cgttgtccag ctgctgagcc ttccctttct
     1441 cccggacagc ccacgctacc tgctcttgga gaagcacaac gaggcaagag ctgtgaaagc
     1501 cttccaaacg ttcttgggta aagcagacgt ttcccaagag gtagaggagg tcctggctga
     1561 gagccgcgtg cagaggagca tccgcctggt gtccgtgctg gagctgctga gagctcccta
     1621 cgtccgctgg caggtggtca ccgtgattgt caccatggcc tgctaccagc tctgtggcct
     1681 caatgcaatt tggttctata ccaacagcat ctttggaaaa gctgggatcc ctctggcaaa
     1741 gatcccatac gtcaccttga gtacaggggg catcgagact ttggctgccg tcttctctgg
     1801 tttggtcatt gagcacctgg gacggagacc cctcctcatt ggtggctttg ggctcatggg
     1861 cctcttcttt gggaccctca ccatcacgct gaccctgcag gaccacgccc cctgggtccc
     1921 ctacctgagt atcgtgggca ttctggccat catcgcctct ttctgcagtg ggccaggtgg
     1981 catcccgttc atcttgactg gtgagttctt ccagcaatct cagcggccgg ctgccttcat
     2041 cattgcaggc accgtcaact ggctctccaa ctttgctgtt gggctcctct tcccattcat
     2101 tcagaaaagt ctggacacct actgtttcct agtctttgct acaatttgta tcacaggtgc
     2161 tatctacctg tattttgtgc tgcctgagac caaaaacaga acctatgcag aaatcagcca
     2221 ggcattttcc aaaaggaaca aagcataccc accagaagag aaaatcgact cagctgtcac
     2281 tgatggtaag ataaatggaa ggccttaact cgaggggggg cccgcggttc gaaggtaagc
     2341 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttgga
     2401 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2461 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2521 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2581 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2641 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2701 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     2761 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     2821 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagcc tttgtctcaa
     2881 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     2941 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     3001 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     3061 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     3121 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     3181 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     3241 tatgtgtggg agggctaagc acaattcgag ctcggtacct ttaagaccaa tgacttacaa
     3301 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     3361 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     3421 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     3481 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     3541 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     3601 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     3661 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3721 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3781 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3841 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3901 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca
     3961 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     4021 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4081 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4141 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4201 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4261 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4321 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4381 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4441 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4501 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4561 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     4621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4981 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5161 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5281 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5641 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5701 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5881 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6481 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6601 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6661 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6721 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
     6781 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca
     6841 atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag
     6901 aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga
     6961 aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat
     7021 attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga
     7081 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7141 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7201 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     7261 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     7321 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     7381 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     7441 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     7501 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     7561 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     7621 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     7681 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     7741 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     7801 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     7861 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     7921 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     7981 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     8041 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     8101 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     8161 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     8221 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     8281 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     8341 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     8401 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     8461 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     8521 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     8581 gacagatcca ttcgatta
//
