LOCUS       sequence-337147-        9367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase Reporter for Human PTEN Enhancer PE - Reverse 
            Orientation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9367)
  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 9367)
  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-hPE(-)-Luc
FEATURES             Location/Qualifiers
     source          1..9367
                     /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"
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 ggtaccctgt caaaaaaaga aaacaacaat
     5041 aacaaaaaaa aacccagcac agttctgtta ggagacatac ctctcctttt ctggctccaa
     5101 aagagaatga atgctttttg aatcctagga ggtaaaccaa tatcgtaatt tcctacatgc
     5161 tacaaaatag ccatctttag taaagctcca atttcagggt ggattttaat tgtttactac
     5221 taataaacct tgggagttaa tttataccat ccctcaagtc ccaacacaaa cagattccct
     5281 tttccagaac tcatggtggc aagattccat agaactcccc agatgatggc agctactatc
     5341 agagctctca actctgacag gaattaacaa ataacaggtt acaaacagga aacacagtgt
     5401 ctcatatcca tggctccact gatgcatggt actatgtaac atgtagggta tgttaattcc
     5461 ctgatattta ttcacttatt cattcgacac atagttattg agtatctcct gtgtaccacg
     5521 ttgtaagctc tggggataca gtgatgaggg caattgacct gatccttgtc ttaatggagt
     5581 ttgattacta gtagggggaa gtcatataaa aagacgatga taaacaagga atcaaataaa
     5641 aacataagta atttcagttt gagaggtcac agaaagttaa taaaaaggtt ctatgagaat
     5701 gaagggaact ctcacacagc ttgtgagagt aggactgtct gtgtagatag catgagatag
     5761 gagaaagagc tagccatgtg ataacatgga gacagggacc agcaaaatat ttaagtctct
     5821 tcagctggaa agactgtgct gtgttctgat aacaaaaagg aggcctccat agctagaaca
     5881 gtgagtccta ggcagagtga ggcaaattaa actggggcca gatcctggga aggattttga
     5941 attattctca ggcatgtttt aagcaggatg gcatagtcca gagaatggat ttcaagggaa
     6001 ggaggcaaga gctggggaag acagggtgcc agaaagtatt gaaaccaaat agcaagttat
     6061 tgcagtggct cagaaggaga aaacaatggt gattatggag atgaacagag tagccttatt
     6121 caaggtgtgt tttagaagta gaatctacag gacttgctga taaattaaac atggaaagaa
     6181 aattaagaca aagatgactt acaggttctt agcttgagca actgggtgaa tggttatgtt
     6241 gtcgactaag cctgaggaag gaacagcttt caaggataga tggaaggaat aattaggtat
     6301 ccaacttcta cgtgttaaat gaaaggcgac tgtgatacac cccagggaaa tatattcagg
     6361 actgccctct gagccctcta gacataggtt tcatttgtac cacaaccatc tccccttcca
     6421 aatacccaca gagatgtttc ctatatctat gttttcaagg tgtgtggtcg tgaagaacag
     6481 ggagcccttc actcctgtct gcattgcccg ctaaactaca ggctgacgtg ccttctgatt
     6541 ttcttttctg ctgtcagagc atttgcttcc ctctgtccta caggacaact ctaaatcatt
     6601 atctccacaa cacctgctcc atatcaaaag tgttagattt ttagccagaa gacatgcact
     6661 atttttctta ctagctgggt gacctggggc atgccattta acttctggga gccctgcagg
     6721 tctttgtaaa gacaatatca atgattacat gtaaaagcac tttgtaaact gtaagcagtt
     6781 acgtaaatag gcttgttatt ctcatttttg tctcagtctc caccactgta acaaataatc
     6841 ttcctttttt tttttttttt tttttgctct gttctcctcg cttttcttct tgcgtgcttc
     6901 ttctgtctct tgtcctttct gccacataaa tttgctttcc tgaaaaaggc tggttatttg
     6961 gagacatcat caccagccaa tcagagctct ctattccttc tggctggaaa gccaatcaga
     7021 tgtaagctcc aggtctcttt tcagtgttgc tagggcttca ggtctacttc taaaagctga
     7081 cttgcttgct ttccgctagt tcattctctg tctctctctg ctccccactt ctcccacccc
     7141 cactctattc ctctcactcc tcccctcttc tgggtttgga gcagttccaa atacttaagg
     7201 cttccggaag tttgggttat ggataagcaa ggtttgactg tatatttttt cccttgagaa
     7261 atagaggaca gctattccat gttcaatatt gaagcaaatg ttttagcagc ttaatgtccc
     7321 ttggctacat aaaatttagt tctgtttatg tgttttgggt gtacagatca ctttttattc
     7381 agatggacta gagcataaac tggcattgag ttttctggca atgaaacgtc atagcactgg
     7441 aacctcactg ggctttacaa aaatgcattc aaaatatttt aaagtacttc cactttataa
     7501 ctgatgacag aagatgataa atacattgtg gttattttgc tcctgttttt tgaactttca
     7561 gtgctgggat tattggagac aactaccctg agatagcagt gatttgggaa ctaataccta
     7621 atggctaagt gggtgtcctc agccctcaca tcctgcttat ctatggaaac taagtaacat
     7681 gggaatgata gctagaaatc cacccttttc agaggaccca agagtccatc tttaaatgct
     7741 gtatggctgg gtatgctgtg aggattctct taacacagaa cagctctgcc ttccaaaacc
     7801 ctaaagacgt ttgataagga aaaatagcta acaagttatg cctttctcat aacctttgaa
     7861 acctacttaa aaactgaaac agtacaagaa tttgcagttt aaaatgacca gagaagtggc
     7921 agaaagccag aattctgtca gagacggctt agaaaagctt aatgattaac aaatgggtca
     7981 ctgccttcag gctctgacct ttagaactaa ccaattttag ccttttggtt atgttccttc
     8041 aactcgtttt atatcaagga aaaaaaaata ctctctaatg gcacagtacc acccatccaa
     8101 aaaaaggatt cttagcagtc cagtatttgg tcagtagctt ttctgataac atggagttgg
     8161 tcctaagcac agtattaaca actgttaaat tatatttttt gttagagttt aaggaaggcc
     8221 ctaagcacga ctgcatatcc tgcaggtgaa atgtacgaaa tttcggttaa catggtaacg
     8281 gctctaaaaa tacctaggct acagtttaac cacagagttt cctccttaac gtaaaaaagc
     8341 caccatcacg agctgttaga aagtgcaaca aaagcaatct ctgtcggact ttgtgcaagc
     8401 aggctattaa atcaggcctg gcagtgacaa tctgcagcca ctctgcatca aagtaataat
     8461 taaagttttg caagcccaaa cagcaatgca caaaccacct aagaaatatg ttacattgat
     8521 aagtcctttt tttctgcctg taacttctct tctcccctac ccccaaccac caggtggact
     8581 aagcagggcg tggcaggggc accgtgacaa agagtcacca caccataacc acagggcccc
     8641 acccgaagga aaacctaggg caggctgcca cggttttccc agcaacctct gcaccaggat
     8701 cccatagagt atgcacagga ggctaaatca ctcacctggg gtttcccatc tgggctcttt
     8761 gcttggcaga gacagagagg gtggggtcaa ccttttcctt gatgcttttg cagttagcaa
     8821 gttggaaaca catttgactc ttttgaattc tgtgccttgg ctattcagga gtctgtcatg
     8881 tctgggctta ctaaattcta ccatgtctag gctgccgggt ttaataaata gaaatacctg
     8941 acatccagtt aaattggaat tttgaataaa cagtgaagaa tttagtgagt gtaagtatat
     9001 cccatgcaat gcttgggata cttacactac atacttatat acttatacta caaataattt
     9061 catagtttac ctgaaattca aatttgactg gatgtacttt cagagtcttt ttatccacca
     9121 acacccaatt aacaaattgg tttttatttg tttgggcttt ttaacttcat tttgctttta
     9181 tttagttttg ctttgccttg caagccttgg gtcagtgtag aaaggctaat ccaaggaaat
     9241 gagctagcat ttgctgagct cattcttaaa tcaggcttca tcttaggtac ttttgatgca
     9301 tggtctttta aaatcctcac atcatcctct ataatagcct acttaaaatc cacattttat
     9361 agaggag
//
