LOCUS       sequence-204620-        7899 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A lentiviral vector for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7899)
  AUTHORS   Brown FC, Still E, Koche RP, Yim CY, Takao S, Cifani P, Reed C, 
            Gunasekera S, Ficarro SB, Romanienko P, Mark W, McCarthy C, de 
            Stanchina E, Gonen M, Seshan V, Bhola P, O'Donnell C, Spitzer B, 
            Stutzke C, Lavallee VP, Hebert J, Krivtsov AV, Melnick A, Paietta 
            EM, Tallman MS, Letai A, Sauvageau G, Pouliot G, Levine R, Marto JA,
            Armstrong SA, Kentsis A
  TITLE     MEF2C Phosphorylation Is Required for Chemotherapy Resistance in 
            Acute Myeloid Leukemia.
  JOURNAL   Cancer Discov. 2018 Apr;8(4):478-497. doi: 
            10.1158/2159-8290.CD-17-1271. Epub 2018 Feb 5.
  PUBMED    29431698
REFERENCE   2  (bases 1 to 7899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1158/2159-8290.CD-17-1271"; journalName: "Cancer Discov"; date: 
            "2018-04"; volume: "8"; issue: "4"; pages: "478-497"
COMMENT     Sequence Label: pRecLV105
FEATURES             Location/Qualifiers
     source          1..7899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2..26)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    101..218
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        267..777
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     696..714
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             799..1398
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(799..818)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1295..1315
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1526..1759
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1831..1952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1868..1887)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1922..1941
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1992..2127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2147..2166)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2148..2166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2176..2193)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2176..2192)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2185..2207)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2334..2789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2421..2440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2631..2652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2815..2919
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2920..2988,2989..3780)
                     /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(3138..3157)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3951..4539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4440..4459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4693..4710
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4827..4848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4863..4868,4869..4886,4887..4893)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4901..4917
                     /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     4906..4928
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4925..4941
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4925..4941
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4960..4980
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4962..4980
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5008..5234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5235..5415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5462..5587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6080..6313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6498..6542
                     /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"
     promoter        6840..7088
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6840..6860
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7011..7030
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        7243..7622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7623..7826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7777..7797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7823..7847
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aacccagctt tcttgtacaa agtggttgat cgcgtgcatg cgacgtcata gctctctccc
       61 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
      121 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      181 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      241 gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct
      301 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
      361 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
      421 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
      481 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
      541 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
      601 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
      661 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
      721 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
      781 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
      841 gacgtcccca gggcggtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
      901 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
      961 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     1021 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     1081 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     1141 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     1201 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     1261 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     1321 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     1381 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     1441 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     1501 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     1561 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     1621 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     1681 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     1741 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     1801 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     1861 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1921 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     1981 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2041 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2101 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     2161 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2221 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2281 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     2341 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2401 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2461 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2521 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2581 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2641 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2701 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2761 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     2821 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2881 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2941 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3001 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3061 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3121 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3181 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3241 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3301 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3361 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3421 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3481 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3541 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3601 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3661 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3721 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3781 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3841 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3901 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3961 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4021 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4081 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4141 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4201 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4261 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4321 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4381 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4441 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4501 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4561 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4621 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4681 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4741 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4801 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4861 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4921 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     4981 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     5041 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     5101 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     5161 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     5221 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5281 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5341 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5401 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     5461 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     5521 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     5581 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     5641 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     5701 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     5761 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     5821 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     5881 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     5941 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     6001 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     6061 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     6121 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     6181 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     6241 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     6301 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     6361 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     6421 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     6481 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     6541 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     6601 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     6661 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     6721 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     6781 tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg
     6841 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     6901 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     6961 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     7021 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     7081 gaaacaccgg tatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
     7141 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
     7201 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
     7261 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     7321 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     7381 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     7441 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     7501 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     7561 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     7621 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     7681 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     7741 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     7801 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     7861 atccacgctg ttttgacctc catagaagat tctagcgcg
//
