LOCUS       sequence-202269-        7989 bp    DNA     circular SYN 15-DEC-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 7989)
  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 7989)
  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"; 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..7989
                     /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             1537..1770
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1842..1963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1879..1898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1933..1952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2003..2138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2158..2177)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2159..2177)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2187..2204)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2187..2203)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2196..2218)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2345..2800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2432..2451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2642..2663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2826..2930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2931..2999,3000..3791)
                     /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(3149..3168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3962..4550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4451..4470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4917..4937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4952..4957,4958..4975,4976..4982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4990..5006
                     /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     4995..5017
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5014..5030
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5014..5030
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5049..5069
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5051..5069
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5097..5323
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5324..5504
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5551..5676
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6169..6402
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6587..6631
                     /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        6929..7177
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6929..6949
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7100..7119
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        7333..7712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7713..7916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7867..7887
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7913..7937
                     /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 gagatccagt 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 catcgttaag agctcggtac ctttaagacc aatgacttac aaggcagctg
     1501 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     1561 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     1621 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     1681 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1741 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     1801 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     1861 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1921 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     1981 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2041 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2101 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     2161 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     2221 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2281 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2341 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2401 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2461 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2521 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2581 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2641 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2701 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2761 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2821 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2881 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2941 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3001 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3061 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3121 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3181 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3241 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3301 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3361 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3421 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3481 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3541 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3601 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3661 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3721 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3781 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3841 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3901 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3961 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4021 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4081 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4141 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4201 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4261 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4321 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4381 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4441 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4501 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4561 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4621 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4681 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgccctt
     4741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4801 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4861 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaaaatg
     4921 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     4981 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     5041 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
     5101 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
     5161 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
     5221 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
     5281 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
     5341 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5401 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5461 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     5521 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     5581 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     5641 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     5701 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     5761 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     5821 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     5881 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     5941 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     6001 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     6061 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     6121 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     6181 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     6241 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     6301 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     6361 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     6421 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     6481 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     6541 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     6601 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     6661 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     6721 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     6781 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     6841 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gatcacgaga
     6901 ctagcctcga gcggccgccc ccttcaccga gggcctattt cccatgattc cttcatattt
     6961 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
     7021 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     7081 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     7141 cttggcttta tatatcttgt ggaaaggacg aaacaccggt gatctataca ttgaatcaat
     7201 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     7261 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     7321 gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     7381 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     7441 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     7501 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     7561 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
     7621 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
     7681 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     7741 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     7801 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     7861 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     7921 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagat
     7981 tctagcgcg
//
