LOCUS       sequence-207773-        8002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gamma-Retroviral transfer vector for expressing HuR (wild type), 
            IRES-driven Puromycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8002)
  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  JOURNAL   Unpublished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 8002)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pQC V5 HuR.wt IRES Puro
FEATURES             Location/Qualifiers
     source          1..8002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             514..689
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    752..951
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1152..1568
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1498..1520
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1536..1552
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1605..1908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1909..2112
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2109..2133
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2267..2308
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3331..3903
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3498..3515)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3725..3744
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3940..4539
                     /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(3940..3959)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4436..4456
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4708..5133
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5236..5258)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5394..5414)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5399..5728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5579..5714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5750..5769
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5850..5867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6021..6609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6101..6120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6780..7571,7572..7640))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7403..7422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7641..7745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7813..7831
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7931..8002,1..309)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgctctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact
      541 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
      601 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
      661 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
      721 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
      781 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
      841 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      901 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      961 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     1021 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     1081 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1141 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1201 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1261 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1321 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1381 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1441 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1501 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1561 taggcgccgg aattgaagat ctgggggatc gatcctctag agtccgttac ataacttacg
     1621 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1681 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1741 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1801 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1861 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1921 tggcagtaca tcaatgggcg tgaatagcgg tttgactcac ggggatttcc aagtctccac
     1981 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2041 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2101 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2161 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga
     2221 acggacctgc aggaattgat ccgcggccgc accggggatc accatgggta agcctatccc
     2281 taaccctctc ctcggtctcg attctacggg atccggttct agagccaccg gtatgtctaa
     2341 tggttatgaa gaccacatgg ccgaagactg caggggtgac atcgggagaa cgaatttgat
     2401 cgtcaactac ctccctcaga acatgaccca ggatgagtta cgaagcctgt tcagcagcat
     2461 tggtgaagtt gaatctgcaa aacttattcg ggataaagta gcaggacaca gcttgggcta
     2521 tggctttgtg aactacgtga ccgcgaagga tgcagagaga gcgatcaaca cgctgaacgg
     2581 cttgaggctc cagtcaaaaa ccattaaggt gtcgtatgct cgcccgagct cagaggtgat
     2641 caaagacgcc aacttgtaca tcagcgggct cccgcggacc atgacccaga aggacgtaga
     2701 agacatgttc tctcggtttg ggcggatcat caactcgcgg gtcctcgtgg atcagactac
     2761 aggtttgtcc agaggggttg cgtttatccg gtttgacaaa cggtcggagg cagaagaggc
     2821 aattaccagt ttcaatggtc ataaaccccc aggttcctct gagcccatca cagtgaagtt
     2881 tgcagccaac cccaaccaga acaaaaacgt ggcactcctc tcgcagctgt accactcgcc
     2941 agcgcgacgg ttcggaggcc ccgttcacca ccaggcgcag agattcaggt tctcccccat
     3001 gggcgtcgat cacatgagcg ggctctctgg cgtcaacgtg ccaggaaacg cctcctccgg
     3061 ctggtgcatt ttcatctaca acctggggca ggatgccgac gaggggatcc tctggcagat
     3121 gtttgggccg tttggtgccg tcaccaatgt gaaagtgatc cgcgacttca acaccaacaa
     3181 gtgcaaaggg tttggctttg tgaccatgac aaactatgaa gaagccgcga tggccatagc
     3241 cagcctgaac ggctaccgcc tgggggacaa aatcttacag gtttccttca aaaccaacaa
     3301 gtcccacaaa tagaattctg attaattccg cccctctccc tccccccccc ctaacgttac
     3361 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     3421 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     3481 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     3541 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     3601 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     3661 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     3721 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     3781 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     3841 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3901 taagcttgat cacgcgtctc gagatcaccg gagcttacca tgaccgagta caagcccacg
     3961 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4021 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4081 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4141 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4201 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4261 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4321 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4381 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4441 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4501 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     4561 cgcccgaccg aaaggagcgc acgaccccat gcatcgatca gtggtccagg ctctagtttt
     4621 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     4681 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     4741 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt
     4801 cagatcaagg tcaggaacag atggaacagg gtcgacccta gagaaccatc agatgtttcc
     4861 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4921 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     4981 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5041 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5101 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     5161 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg
     5221 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5281 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5341 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tagatccggc tgtggaatgt
     5401 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5461 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5521 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5581 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5641 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5701 cttttttgga ggcctaggct tttgcaaaaa gcttactggc ttaactatgc ggcatcagag
     5761 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     5821 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5881 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6421 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6601 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6661 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6721 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6781 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6841 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6901 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6961 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7021 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7081 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7141 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7201 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7261 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7321 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7381 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7441 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7501 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7561 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7621 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7681 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7741 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7801 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt agcttggcca
     7861 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta
     7921 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     7981 gttcatagcc catatatgga gt
//
