LOCUS       sequence-207837-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression plasmid of cDNA with neomycin resistance gene 
            (empty vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7210)
  AUTHORS   Tarumoto Y, Lu B, Somerville TDD, Huang YH, Milazzo JP, Wu XS, 
            Klingbeil O, El Demerdash O, Shi J, Vakoc CR
  TITLE     LKB1, Salt-Inducible Kinases, and MEF2C Are Linked Dependencies in 
            Acute Myeloid Leukemia.
  JOURNAL   Mol Cell. 2018 Feb 28. pii: S1097-2765(18)30109-6. doi: 
            10.1016/j.molcel.2018.02.011.
  PUBMED    29526696
REFERENCE   2  (bases 1 to 7210)
  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"; journalName: "Mol Cell. 
            2018 Feb 28. pii: S1097-2765(18)30109-6. doi: 
            10.1016/j.molcel.2018.02.011."
COMMENT     Sequence Label: LentiV_Neo
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..248
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             249..429
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    476..601
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1094..1327
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1512..1556
                     /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"
     misc_feature    1740..1857
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1926..2137
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2157..2166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2166..2189
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2199..2255
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2274..3068
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2328..2347)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2938..2957
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    3084..3672
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3137..3157)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3555..3566)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3744..3977
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4055..4176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4092..4111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4146..4165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4216..4351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4371..4390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4372..4390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4400..4417)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4400..4416)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4409..4431)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4558..5013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4645..4664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4855..4876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5039..5143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5144..5212,5213..6004)
                     /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(5362..5381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6175..6763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6664..6683
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6917..6934
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7051..7072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7087..7092,7093..7110,7111..7117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7125..7141
                     /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     7130..7152
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7149..7165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7149..7165
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7184..7204
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7186..7204
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg
       61 taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg
      121 tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga
      181 ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat
      241 acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
      301 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
      361 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
      421 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
      481 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
      541 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
      601 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
      661 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
      721 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
      781 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
      841 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
      901 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
      961 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1021 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1081 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1141 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1201 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1261 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1321 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     1381 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     1441 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     1501 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     1561 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     1621 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     1681 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc agaccacaat
     1741 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     1801 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     1861 gtttattaca gggacagcag agatccactt tggcgccggc tcgagtggct ccggtgcccg
     1921 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     1981 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2041 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2101 cgttcttttt cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg ggatccgcca
     2161 ccatggatta caaagacgat gacgataagg gcagcggcgc caccaacttc agcctgctga
     2221 agcaggccgg cgacgtggag gagaaccccg gccccatgac cgagtacaag cccatgattg
     2281 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2341 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2401 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     2461 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2521 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2581 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2641 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2701 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2761 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     2821 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     2881 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     2941 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3001 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3061 tcttctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     3121 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3181 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3241 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3301 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3361 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3421 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3481 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3541 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3601 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3661 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     3721 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     3781 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     3841 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     3901 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     3961 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     4021 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     4081 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4141 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     4201 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4261 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4321 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     4381 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     4441 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     4501 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     4561 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4621 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4681 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4741 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4801 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4861 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4921 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4981 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     5041 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5101 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5161 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5221 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5281 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     5341 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5401 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5461 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5521 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5581 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5641 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5701 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5761 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5821 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5881 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5941 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6001 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6061 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6121 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6181 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6241 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6301 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6361 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6421 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6481 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6541 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6601 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6661 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6721 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6781 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6841 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6901 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     6961 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     7021 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     7081 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     7141 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     7201 aagggaacaa
//
