LOCUS       sequence-380682-        8436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GLUT3 protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8436)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8436)
  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 GLUT3
FEATURES             Location/Qualifiers
     source          1..8436
                     /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      647..656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2172..2213
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2250..2270)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2255..2584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2435..2570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2632..2679
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2698..3096
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3183..3416
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3488..3609
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3525..3544)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3579..3598
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3649..3784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3804..3823)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3805..3823)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3833..3850)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3833..3849)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3842..3864)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3991..4446
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4078..4097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4288..4309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4472..4576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4577..4645,4646..5437)
                     /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(4795..4814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5608..6196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6097..6116
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6350..6367
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6484..6505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6520..6525,6526..6543,6544..6550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6558..6574
                     /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     6563..6585
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6582..6598
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6582..6598
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6617..6637
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6619..6637
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6665..6891
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6892..7072
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7119..7244
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7737..7970
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8155..8199
                     /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 cactagttaa ctgcaggcca ccatggggac
      661 acagaaggtc accccagctc tgatatttgc catcacagtt gctacaatcg gctctttcca
      721 atttggctac aacactgggg tcatcaatgc tcctgagaag atcataaagg aatttatcaa
      781 taaaactttg acggacaagg gaaatgcccc accctctgag gtgctgctca cgtctctctg
      841 gtccttgtct gtggccatat tttccgtcgg gggtatgatc ggctcctttt ccgtcggact
      901 cttcgtcaac cgctttggca ggcgcaattc aatgctgatt gtcaacctgt tggctgtcac
      961 tggtggctgc tttatgggac tgtgtaaagt agctaagtcg gttgaaatgc tgatcctggg
     1021 tcgcttggtt attggcctct tctgcggact ctgcacaggt tttgtgccca tgtacattgg
     1081 agagatctcg cctactgccc tgcggggtgc ctttggcact ctcaaccagc tgggcatcgt
     1141 tgttggaatt ctggtggccc agatctttgg tctggaattc atccttgggt ctgaagagct
     1201 atggccgctg ctactgggtt ttaccatcct tcctgctatc ctacaaagtg cagcccttcc
     1261 attttgccct gaaagtccca gatttttgct cattaacaga aaagaagagg agaatgctaa
     1321 gcagatcctc cagcggttgt ggggcaccca ggatgtatcc caagacatcc aggagatgaa
     1381 agatgagagt gcaaggatgt cacaagaaaa gcaagtcacc gtgctagagc tctttagagt
     1441 gtccagctac cgacagccca tcatcatttc cattgtgctc cagctctctc agcagctctc
     1501 tgggatcaat gctgtgttct attactcaac aggaatcttc aaggatgcag gtgttcaaga
     1561 gcccatctat gccaccatcg gcgcgggtgt ggttaatact atcttcactg tagtttctct
     1621 atttctggtg gaaagggcag gaagaaggac tctgcatatg ataggccttg gagggatggc
     1681 tttttgttcc acgctcatga ctgtttcttt gttattaaag gataactata atgggatgag
     1741 ctttgtctgt attggggcta tcttggtctt tgtagccttc tttgaaattg gaccaggccc
     1801 cattccctgg tttattgtgg ccgaactctt cagccagggc ccccgcccag ctgcgatggc
     1861 agtggccggc tgctccaact ggacctccaa cttcctagtc ggattgctct tcccctccgc
     1921 tgctcactat ttaggagcct acgtttttat tatcttcacc ggcttcctca ttaccttctt
     1981 ggcttttacc ttcttcaaag tccctgagac ccgtggcagg acttttgagg atatcacacg
     2041 ggcctttgaa gggcaggcac acggtgcaga tagatctgga aaggacggcg tcatggagat
     2101 gaacagcatc gagcctgcta aggagaccac caccaatgtc taactcgagt ctagagggcc
     2161 cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg
     2221 gttagtaatg agtttggaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2341 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2401 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2461 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2521 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2581 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     2641 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     2701 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     2761 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     2821 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     2881 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     2941 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     3001 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     3061 gaattgctgc cctctggtta tgtgtgggag ggctaagcac aattcgagct cggtaccttt
     3121 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     3181 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     3241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     3301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     3361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     3421 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     3481 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3541 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3601 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
     3661 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3721 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3781 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     3841 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3901 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3961 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     4021 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4081 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4141 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4201 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4261 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4321 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4381 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4441 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4501 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4561 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4621 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4681 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4741 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4801 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4861 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4921 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4981 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5041 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5101 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5161 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5221 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5281 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5341 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5401 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5461 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5521 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5581 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5641 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5701 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5761 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5821 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5881 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5941 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6001 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6061 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6121 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6181 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6241 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6301 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6361 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6481 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6541 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6601 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     6661 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
     6721 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
     6781 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
     6841 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
     6901 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     6961 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7021 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     7081 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     7141 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     7201 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     7261 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     7321 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     7381 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     7441 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     7501 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     7561 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     7621 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     7681 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     7741 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     7801 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     7861 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     7921 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     7981 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     8041 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     8101 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     8161 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     8221 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     8281 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     8341 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     8401 ggtggagaga gagacagaga cagatccatt cgatta
//
