LOCUS       sequence-350788-        7585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty vector to express overexpression cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7585)
  AUTHORS   Lu B, Klingbeil O, Tarumoto Y, Somerville TDD, Huang YH, Wei Y, Wai 
            DC, Low JKK, Milazzo JP, Wu XS, Cao Z, Yan X, Demerdash OE, Huang G,
            Mackay JP, Kinney JB, Shi J, Vakoc CR
  TITLE     A Transcription Factor Addiction in Leukemia Imposed by the MLL 
            Promoter Sequence.
  JOURNAL   Cancer Cell. 2018 Dec 10;34(6):970-981.e8. doi: 
            10.1016/j.ccell.2018.10.015. Epub 2018 Nov 29.
  PUBMED    30503706
REFERENCE   2  (bases 1 to 7585)
  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: 
            "10.1016/j.ccell.2018.10.015"; journalName: "Cancer Cell"; date: 
            "2018-12-10- 10"; volume: "34"; issue: "6"; pages: "970-981.e8"
COMMENT     Sequence Label: pHAGE_puro
FEATURES             Location/Qualifiers
     source          1..7585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..71
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             140..739
                     /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(140..159)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     636..656
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    753..1341
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(806..826)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1224..1235)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1416..1649
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(1768..1786)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1854..1958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1959..2027,2028..2819)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2177..2196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2990..3578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3479..3498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3732..3749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3989..4010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4025..4030,4031..4048,4049..4055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4063..4079
                     /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     4068..4090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4087..4103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4087..4103
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4153..4172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4364..4997
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5044..5169
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5666..5899
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6084..6128
                     /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    6421..6538
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6590..6893
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6894..7097
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7048..7068
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7094..7118
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        join(7205..7585,1..119)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7240..7263)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
       61 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
      121 cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
      181 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
      241 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
      301 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
      361 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
      421 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
      481 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
      541 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
      601 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
      661 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
      721 ccgcaagccc ggtgcctgaa tcgatagatc ctaatcaacc tctggattac aaaatttgtg
      781 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
      841 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
      901 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
      961 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1021 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     1081 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     1141 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     1201 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     1261 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     1321 ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca aggcagctgt
     1381 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     1441 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     1501 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     1561 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     1621 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     1681 tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa ttaaggaaag
     1741 ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1801 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1861 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1921 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     2221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2821 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2881 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2941 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3001 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3061 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3121 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3601 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3781 ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac taattttttt
     3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3901 cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg tttcccgact
     3961 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4021 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4081 ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca tttttttcac
     4141 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
     4201 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
     4261 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     4321 aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg gctaattcac
     4381 tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     4441 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     4501 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     4561 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
     4621 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
     4681 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
     4741 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
     4801 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4861 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4921 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4981 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     5041 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     5101 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     5161 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     5221 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     5281 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     5341 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     5401 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     5461 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga
     5521 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     5581 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     5641 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     5701 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     5761 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     5821 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     5881 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     5941 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     6001 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     6061 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     6121 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     6181 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     6241 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     6301 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     6361 gattagtgaa cggatctcga cggtatcgcc gaattcacaa atggcagtat tcatccacaa
     6421 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     6481 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     6541 ggtttattac agggacagca gagatccagt ttggactagt ggagttccgc gttacataac
     6601 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     6661 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     6721 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     6781 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     6841 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     6901 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     6961 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     7021 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     7081 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     7141 gttttgacct ccatagaaga caccggcggc cgcaccggtc tgcagctagc tcgagaattc
     7201 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     7261 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     7321 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     7381 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     7441 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     7501 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     7561 aggggtgggt ccgggggcgg gctca
//
