LOCUS       sequence-337072-        9077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase Reporter for Mouse PTEN Enhancer PE - Forward 
            Orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9077)
  AUTHORS   Tottone L, Lancho O, Loh JW, Singh A, Kimura S, Roels J, Kuchmiy A, 
            Strubbe S, Lawlor MA, da Silva-Diz V, Luo S, Gachet S, Garcia-Prieto
            CA, Hagelaar R, Esteller M, Meijerink JPP, Soulier J, Taghon T, Van 
            Vlierberghe P, Mullighan CG, Khiabanian H, Rocha PP, Herranz D
  TITLE     A Tumor Suppressor Enhancer of PTEN in T-cell development and 
            leukemia.
  JOURNAL   Blood Cancer Discov. 2021 Jan;2(1):92-109. doi: 
            10.1158/2643-3230.BCD-20-0201. Epub 2020 Nov 24.
  PUBMED    33458694
REFERENCE   2  (bases 1 to 9077)
  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: 
            "10.1158/2643-3230.BCD-20-0201"; journalName: "Blood Cancer Discov";
            date: "2021-01"; volume: "2"; issue: "1"; pages: "92-109"
COMMENT     Sequence Label: pGL3-Promoter-mPE(+)-Luc
FEATURES             Location/Qualifiers
     source          1..9077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      274..283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             280..1932
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(346..364)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1973..2094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1984..2003)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2038..2057
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2342..2359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2513..3101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2593..2612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3272..4063,4064..4132))
                     /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     3895..3914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4133..4237)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4264..4719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4351..4370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4561..4582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4850..4898
                     /note="synthetic polyadenylation signal"
     misc_feature    4912..5003
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4952..4971
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             7998..8009
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
      301 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
      361 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
      421 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
      481 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
      541 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
      601 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
      661 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
      721 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
      781 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
      841 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
      901 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
      961 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     1021 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     1081 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     1141 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     1201 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     1261 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     1321 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     1381 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     1441 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     1501 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     1561 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     1621 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     1681 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     1741 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     1801 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     1861 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     1921 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     1981 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2041 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2101 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2161 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     2221 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2281 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     2341 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2401 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2461 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2521 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2581 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2641 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2701 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2761 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2821 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2881 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2941 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3001 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3061 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3361 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3421 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3541 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4201 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4261 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4681 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     4741 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     4801 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc
     4861 tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc
     4921 tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag
     4981 tgcaggtgcc agaacatttc tctatcgata ggtaccgcat ggagatattt cttgagcgat
     5041 gatttcaacc tctgtgtagc aaagggcagt gagccctacg ccctttctgc atctcccact
     5101 aaactgtagc tgatgtggct gctggtggtc ttttctgtga tcagaggatt agctcaattc
     5161 ctcaggataa ctccaaatta ttctctccag agactctgtg gtgtcaacag ccctggatta
     5221 gagaccggtg acctcagctt ttttcttact cgctgtgtaa cctggggaat gtcatttcac
     5281 ttctgcaatc cttgaaggtc cttctgaaga cagtgtcagt aatttcaggg agcctagttt
     5341 gtaaaaactg taagcagcta cataaatagt gattcttgtt ctcattgtcg tctcagctgc
     5401 cacctttgtt acaaataacc tgtcattcat tttgccctgt tcgcttccct ttgcttcttg
     5461 cctgcttctt ctacctcctg tcctttctgc cagataaatt tgcttccctg caaaaggctg
     5521 attatttgga gacatcatca ctagccaatc agagctcagc attccggctg gctggtaagc
     5581 caatcagatg caagctccag gtcttctttc agtgttgcta gggtttcagg tctacttcta
     5641 aaaactggct tgcttgctgc tggttcattc tgtccttctc tccctccatc cccccccccc
     5701 cccatcccct tgctctccaa acccttccct ttccctcctc ccctcttttg ggtttggagc
     5761 aattccaaat gcttaagcct tccggaagtt tgggttatta tgaataagca aggtttaact
     5821 gtgtattttt tcccttgaga aacaggacag ctattccaca ttcgattttg aaagcatgtg
     5881 ttttagcagc ttaatgtccc ttggctccat aaaacagaat ttcgttgtgt ttaagagaat
     5941 ttttttggtg cagagtgact tttattcaga tgactagaag ataaactggc atagaacttg
     6001 gcagtgacgt gtcctgtcac ttgaacctca ttgggctaac acagcgaatt ctaagtatgt
     6061 caaaactcat ccagtttaac tgatgatgga agataatatc tgatagcacc tcagatatta
     6121 gttctatctt gaactttcag tgctgggatt agtggtgaaa atcactggtc agtagtgatg
     6181 gggactctca ctttattgct aatgggtatc ccttccatca catctggctc atctgtggac
     6241 ccatgggtct tctttaaatg ctctctagta gggcgtgtga agatgcccta aacacaaact
     6301 ggctctgtgc tctgaaagct taggggcgat gccggattga gaaggaaaac aacagcaaca
     6361 caagttctgt ctttctcata acctttgcta ctgacttaaa aactggaaca gcataaggat
     6421 tcgcagtgta aaatgaccca agacaggtgg cgcaaagcca gaagtctgtc agaagcagct
     6481 taggaaagct taatgattca caaaggagtc gctgccttca ggctctgacc tttagaacta
     6541 accagcttta agcttttggt tctggccctt caagtcactt gtatcaaaga aaaaggtact
     6601 cccttaaaga cattaccatc catccattca aaaagaaaaa aaatagaaga gaaaaggtgt
     6661 cagcaccagc gactctgttc tcagataaca tggtcttagt cctaaagaca gtattaattg
     6721 cattaattgc ttaaatacat tttttttttt tggttagctt ttagggaagg ccttggtgtt
     6781 actgcatatc ctgcaggtga aatgtatgaa atttggttaa catggtaacg gctctaaaaa
     6841 tacctaggct acagtttaac cacagagttt cctccttaac gtaaaagagc cgctgtcatg
     6901 aggtgtacga aagtgcaaca aaggcgatcc gtctgacttc atgcaagcag cctattaaga
     6961 caagcctggc gaggacactg tgcggccact aggcatcaaa gcaggaatta aagttttgtg
     7021 agaccatctg acaaagcaca acccagctga tcaaaacggc taagcattct ttccctgttt
     7081 ctacctcttc tgcccccacc cactggcaac caagcagggt gtggtggggg ttcaaacagt
     7141 cactatagcg tgaccacagg gcctcaccca aggaaaacct gggtcacaga gacccaagcg
     7201 ggtaaggagg ctgcagcagg ctccccagca ttctctgcag caggctcatg tggagggtgc
     7261 agggaaggga aaattgctca cttgaggttt tccccatgag gcccttgctt cacaaaacaa
     7321 agggtggcag agggtgcgac ctttcccagg atggctctgc agttgcccac gtcaggctct
     7381 tctgaactcc gagtgtcctt acagggttaa gagatttctt acaaaaatca ggctgtcgag
     7441 gttgcatggt ttggtaaatg aaaacactga cacccattta tactagaatt tcagataaac
     7501 aatgaataat ttatttagta aaagaatgtc tggtgcaata tttgggatac tgaagaaatc
     7561 cattgttcat ctgaaattca gatctgggtg ttgtgtgttc tctctgatag ctctagtcaa
     7621 atccttccat ccttcgtcct taacttctca ccttcgagtg cccaaacaac atgctcaact
     7681 gtactttttg cttcaggtat ttagctttgg cttgttctgg ctttgtttgg tcttgtttca
     7741 ccctgcctct taaacctccg ctcattatac agagccagag gtgaggagat gaagtagcag
     7801 ctgagctccg tctcaccttt gtgttatgga cttttaatgc atgtaatttt tttaatattt
     7861 cattttatgt gtatgacttg cctgcatgtg tgcctgtgca gcatgtatgt gtctggtgcc
     7921 tgccggggcc cgaggagggc atcagacccc ctggaacagg agttgggtgg ttgtaagagg
     7981 tctggtcagg cctgggaata gaaggtaggt ccgctgaaac agccattgac cttacctact
     8041 gagccgtctc tcctgcccaa tgaatggtct ttttggaggg gtatcgtctc tgtgtaacct
     8101 tggctgtcct ggaacttgtc agcttgacct cgaactcaca gagatgtgtt tgcctctgcc
     8161 tcccaagtac tgggattaaa ggtgtgcatc agcttaggct gagtggtctt tttcatcatc
     8221 gtaacattct gtgtgatatg ctacttagag cccacatgtt atggataagg ttgctgggct
     8281 cagaaagagt acatgactcg accaaggtca tacagttagc aagtaccaaa gccaagggag
     8341 aatgatgcta agctgttgac tcagttacct agtccacagc atcagtcctg tttaaatgtg
     8401 gctttagtgg ttaattcatg ttaggttctc ttttataaca aatggctcgt ggctcataag
     8461 tgaacgacac aacttgccag ctggattggg actgttttcc ctcagcttgg actaaactac
     8521 tatgtttgtt ttgtgtgtaa gtgcacctcc aacgtacttc tgattgctct tccctagagt
     8581 agctctaagt gctctctgca tgctatgtgt tgtgatgtat acgatgtgtc catgtggtct
     8641 cttaagtggt actgaggctg taggaacatc tgtaactgat tggaaatttc caactgacag
     8701 gttaaaaagc cacggctcct aggtgtaaag ccatttattg aaagccacac atcttatact
     8761 tggcagctaa gaagtccata tttaagtgga agttgattga cttatcagtg tacaactttg
     8821 ggaaagactc tgaacttcca tgtctctact agataattat aaatgggcaa ttatatatga
     8881 aagcttctct catatagagg aagagctggc cacatggtga gcactcaaca aagactgaga
     8941 tagtggtttt ggaaatgatc ttctactggg tattatagtt tggatataaa atagttccaa
     9001 atgcctttct gatgaaggct tagtctccat atagtattgc cttgaggagt gattggatca
     9061 ggaagtgtca acatcac
//
