LOCUS       sequence-207791-        8673 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Luminescence-based glutaminase GAC isoform 3'UTR-mediated RNA 
            stability reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8673)
  AUTHORS   Sandra Martha Gomes Dias
  TITLE     HuR (ELAVL1) silencing sensitizes triple-negative breast cancer to 
            glutaminase-targeted therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: psiCHECK2 3'UTR GAC
FEATURES             Location/Qualifiers
     source          1..8673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    4088..4136
                     /note="synthetic polyadenylation signal"
     promoter        4145..4896
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      4926..4935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4932..6584
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             5891..5899
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    6628..6749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6639..6658)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6693..6712
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6882..6986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6987..7055,7056..7847)
                     /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(7205..7224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8018..8606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8507..8526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagaa atgggttcta gtttcagaat gtttcttcat
     1681 ttaatctttc aaacatcttt agcttttttt tgcaagttat aaatatttat ttgaggtatt
     1741 ttttgttctc aatcttgggt gctggagcca taaagctttt ttttcctttt aatctttgta
     1801 taaaggcagt agattaagaa gtgcatttgt tggtctttaa aaagtattta caagtacata
     1861 aatttgcttt atttttaaaa atacaaaaag gaaaaattta aatttttttt gatgtaatta
     1921 aaatgttaac tatgtggtca gataatccca ttttacaata gtaacagaaa attgtaattc
     1981 ttagttctaa aattcacaaa ttaaactcat aagttttgtt gcattttgtt ttttcttttc
     2041 catttttaaa actaatgtga tgtctttagt ggcaatagaa ggtacttcta tgctaaatac
     2101 aaaactaaaa aggcaaaata atgaacccca aattatttta tttaaaatag cagtggatta
     2161 taaaattagc ttgtgtttac atttatgcca tttttggtga tagattggct ttacatttta
     2221 aaaaatttat ttaaaaattt atcaaatgct ttaaaatatg actcctactt tttttatttt
     2281 gcaactcctc tgttctgtca gagttgttat atacaggagt gtcttatgtt actaaaacat
     2341 tccagccaaa gaatttcaga tgtgagataa tgatgtttca tcaataaaaa gctataatgg
     2401 ttagttactc agaaggagaa acagtgagtg tcttcaagtg aattgttcac ctaaacaatt
     2461 ttattttcat attatccaca taactttttc tatgttatat ttaaatatga atggcaaatt
     2521 ttggttttta gcttttacat tttattatct taattttata aatgctaata tttcttttgt
     2581 gataagttat agcatctcat aaagtttgtt ctatttgaag ttttttagag tacttgagaa
     2641 atgaatttag tctgcaggta gtaagtatgc tactaaaata cgttagatct aaatcctttt
     2701 atttggtata aaaatgcaat attgagaatc aaaacttgtt tttaagagaa ctatagattc
     2761 tacacaacct gatttcaagt aattattcat agtatttata gttgtcttgg caaagtgatt
     2821 gtaaaattct gtaggaccta ttcacacttc ttccttcttc catatacttc tctggttttc
     2881 cccatagttc ccctataatt tcaagtttgt tgaaacctgt taattttagt gggggattag
     2941 aagaaaaact tggtggtttc ttagcatgat ggtgtatgta tgtggtaatg gaaagtctgt
     3001 aaaagtaaat atagtgtagc aaaaaagatt tcactgagta ttttagatac tagtgcaaat
     3061 aaagatagaa aatcttgatc ataatgtctt aagtttggga actgtgatat taagaaaaga
     3121 aattcccttc tagaggtgct ggccaaaaag ccttttgggc taacttaagt attaaattta
     3181 tatatttaaa taattatatt ttaagttgta gaggattttc ccaaggattt tatgcttact
     3241 tgaatgttct ttgaatgttc agatgcatat cctaactgga tgcttctcaa ggccttactg
     3301 catatttgtg ttgcatattt atgttagttg caccagggcc atttgtagtt tgggcaaccg
     3361 aatgccttaa ttggaaaaaa ggcattgtgg tttcccctat gatctaaatt gttacatttt
     3421 accatttcat tccgaagttg gttttacttt attaaatgaa gatttagttt tcatatcgta
     3481 tacatagctg tatagatttc aaaattaggt tgttaatttg tgtcacttac tatttttgtg
     3541 ttggtaatgc tttaaatgca tacttaaaaa tgaagtactg ttatctaagc tactgtgttt
     3601 agaaaatgtt aagaatgagc agaaattttt atagaaaagt ataaacggaa gaagagataa
     3661 gatactgcga ataggccctc aaacttaaaa aagaaaaaac tttgccagtt ttaaggacat
     3721 attttgattc tttcagtatt cttaacacct ttttaaacaa agttcttgat agtacccact
     3781 attattgggt ttgttttatg ccattattga ttcttgatat tcaagcattt acaatgtagc
     3841 atatttgatt ttcttttttc tttctttttt tggcatcatt aacatttcat ttgaaatgca
     3901 tattgttctt gaagtacttt gtttttagca taaatgttgt gcattttatc ttagtgtttg
     3961 gatgaaaaca tttgtgttgt ttagctttca tttgctttgt atatttaata atgtaccttt
     4021 attttccagt atgcctacat tttgtattgc acaataaatt tattttaagc tggcggccgc
     4081 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgagga
     4141 tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac
     4201 gcggggggag ggggaaggaa cgaaacactc tcattcggag gcggctcggg gtttggtctt
     4261 ggtggccacg ggcacgcaga agagcgccgc gatcctctta agcacccccc cgccctccgt
     4321 ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg
     4381 cgccccagag tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg
     4441 gcgacgggct cgctgggtcc taggctccat ggggaccgta tacgtggaca ggctctggag
     4501 catccgcacg actgcggtga tattaccgga gaccttctgc gggacgagcc gggtcacgcg
     4561 gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg gggcacaggt acactatctt
     4621 gtcacccgga ggcgcgaggg actgcaggag cttcagggag tggcgcagct gcttcatccc
     4681 cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt
     4741 tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca
     4801 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     4861 ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc ttggcattcc ggtactgttg
     4921 gtaaagccac catggccgat gctaagaaca ttaagaaggg ccctgctccc ttctaccctc
     4981 tggaggatgg caccgctggc gagcagctgc acaaggccat gaagaggtat gccctggtgc
     5041 ctggcaccat tgccttcacc gatgcccaca ttgaggtgga catcacctat gccgagtact
     5101 tcgagatgtc tgtgcgcctg gccgaggcca tgaagaggta cggcctgaac accaaccacc
     5161 gcatcgtggt gtgctctgag aactctctgc agttcttcat gccagtgctg ggcgccctgt
     5221 tcatcggagt ggccgtggcc cctgctaacg acatttacaa cgagcgcgag ctgctgaaca
     5281 gcatgggcat ttctcagcct accgtggtgt tcgtgtctaa gaagggcctg cagaagatcc
     5341 tgaacgtgca gaagaagctg cctatcatcc agaagatcat catcatggac tctaagaccg
     5401 actaccaggg cttccagagc atgtacacat tcgtgacatc tcatctgcct cctggcttca
     5461 acgagtacga cttcgtgcca gagtctttcg acagggacaa aaccattgcc ctgatcatga
     5521 acagctctgg gtctaccggc ctgcctaagg gcgtggccct gcctcatcgc accgcctgtg
     5581 tgcgcttctc tcacgcccgc gaccctattt tcggcaacca gatcatcccc gacaccgcta
     5641 ttctgagcgt ggtgccattc caccacggct tcggcatgtt caccaccctg ggctacctga
     5701 tttgcggctt tcgggtggtg ctgatgtacc gcttcgagga ggagctgttc ctgcgcagcc
     5761 tgcaagacta caaaattcag tctgccctgc tggtgccaac cctgttcagc ttcttcgcta
     5821 agagcaccct gatcgacaag tacgacctgt ctaacctgca cgagattgcc tctggcggcg
     5881 ccccactgtc taaggaggtg ggcgaagccg tggccaagcg ctttcatctg ccaggcatcc
     5941 gccagggcta cggcctgacc gagacaacca gcgccattct gattacccca gagggcgacg
     6001 acaagcctgg cgccgtgggc aaggtggtgc cattcttcga ggccaaggtg gtggacctgg
     6061 acaccggcaa gaccctggga gtgaaccagc gcggcgagct gtgtgtgcgc ggccctatga
     6121 ttatgtccgg ctacgtgaat aaccctgagg ccacaaacgc cctgatcgac aaggacggct
     6181 ggctgcactc tggcgacatt gcctactggg acgaggacga gcacttcttc atcgtggacc
     6241 gcctgaagtc tctgatcaag tacaagggct accaggtggc cccagccgag ctggagtcta
     6301 tcctgctgca gcaccctaac attttcgacg ccggagtggc cggcctgccc gacgacgatg
     6361 ccggcgagct gcctgccgcc gtcgtcgtgc tggaacacgg caagaccatg accgagaagg
     6421 agatcgtgga ctatgtggcc agccaggtga caaccgccaa gaagctgcgc ggcggagtgg
     6481 tgttcgtgga cgaggtgccc aagggcctga ccggcaagct ggacgcccgc aagatccgcg
     6541 agatcctgat caaggctaag aaaggcggca agatcgccgt gtaataattc tagagtcggg
     6601 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     6661 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     6721 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     6781 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     6841 aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6901 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6961 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7021 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     7081 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     7141 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     7201 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     7261 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     7321 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7381 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7441 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7501 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7561 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7621 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7681 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7741 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7801 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7861 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     8161 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8641 ccttttgctg gccttttgct cacatggctc gac
//
