LOCUS       sequence-227209-        8607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gamma-Retroviral transfer vector for expressing HuR (delHNS), V5 and
            mKO2 tags, IRES-driven Puromycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8607)
  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  JOURNAL   Unpublished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 8607)
  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            "Unpublished"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 mKO2 HuR.DelHNS IRES Puro
FEATURES             Location/Qualifiers
     source          1..8607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             585..760
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    823..1022
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1223..1639
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1569..1591
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1607..1623
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1676..1979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1980..2183
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2180..2204
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2338..2379
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2389..3042
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             2389..3042
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     misc_feature    4114..4576
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4170..4187)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4397..4416
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4617..5216
                     /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(4617..4636)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5113..5133
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5385..5810
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5913..5935)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6071..6091)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6076..6405
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6256..6391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6427..6446
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6527..6544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6698..7286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6778..6797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7457..8248,8249..8317))
                     /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     8080..8099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8318..8422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8490..8508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac
      601 aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc
      661 caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct
      721 cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt
      781 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
      841 tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta
      901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      961 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
     1021 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     1081 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     1141 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1201 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1261 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1321 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1381 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1441 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1501 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1561 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1621 cactccttct ctaggcgccg gaattgaaga tctgggggat cgatcctcta gagtccgtta
     1681 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1741 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1801 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1861 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1921 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1981 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     2041 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     2101 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     2161 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     2221 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgggaac
     2281 ggtgcattgg aacggacctg caggaattga tccgcggccg caccggggat caccatgggt
     2341 aagcctatcc ctaaccctct cctcggtctc gattctacgg gatctaccat ggtgagcgtg
     2401 atcaagcccg agatgaagat gaggtactac atggacggct ccgtcaatgg gcatgagttc
     2461 acaatcgagg gtgagggcac aggcagacct tacgagggac atcaggagat gacactgcgc
     2521 gtcacaatgg ccgagggcgg gccaatgcct ttcgccttcg acctggtgtc ccacgtgttc
     2581 tgttacggcc acagagtttt taccaagtac ccagaagaga tcccagacta tttcaagcag
     2641 gcctttcctg agggcctgtc ctgggagagg tccctggagt tcgaggacgg cggctccgcc
     2701 tccgtgagcg cccacatcag cctgaggggc aacaccttct accacaagtc caagttcacc
     2761 ggcgtgaact tccccgccga cggccccatc atgcagaacc agagcgtgga ctgggagccc
     2821 tccaccgaga agatcaccgc cagcgacggc gtgctgaagg gcgacgtgac catgtacctg
     2881 aagctggagg gcggcggcaa ccacaagtgc cagatgaaga ccacctacaa ggccgccaag
     2941 gagatcctgg agatgcccgg cgaccactac atcggccaca ggctggtgag gaagaccgag
     3001 ggcaacatca ccgagcaggt ggaggacgcc gtggcccact ccggtggcgg aggctcgggc
     3061 ggaggtgggt cgggatccgg ttctagagcc accggtatgt ctaatggtta tgaagaccac
     3121 atggccgaag actgcagggg tgacatcggg agaacgaatt tgatcgtcaa ctacctccct
     3181 cagaacatga cccaggatga gttacgaagc ctgttcagca gcattggtga agttgaatct
     3241 gcaaaactta ttcgggataa agtagcagga cacagcttgg gctatggctt tgtgaactac
     3301 gtgaccgcga aggatgcaga gagagcgatc aacacgctga acggcttgag gctccagtca
     3361 aaaaccatta aggtgtcgta tgctcgcccg agctcagagg tgatcaaaga cgccaacttg
     3421 tacatcagcg ggctcccgcg gaccatgacc cagaagtacg tagaagacat gttctctcgg
     3481 tttgggcgga tcatcaactc gcgggtcctc gtggatcaga ctacaggttt gtccagaggg
     3541 gttgcgttta tccggtttga caaacggtcg gaggcagaag aggcaattac cagtttcaat
     3601 ggtcataaac ccccaggttc ctctgagccc atcacagtga agtttgcagc caaccccaac
     3661 cagaacaaaa acgtggcact cctctcgcag ctgtaccact cgccagcgaa gcttggaaac
     3721 gcctcctccg gctggtgcat tttcatctac aacctggggc aggatgccga cgaggggatc
     3781 ctctggcaga tgtttgggcc gtttggtgcc gtcaccaatg tgaaagtgat ccgcgacttc
     3841 aacaccaaca agtgcaaagg gtttggcttt gtgaccatga caaactatga agaagccgcg
     3901 atggccatag ccagcctgaa cggctaccgc ctgggggaca aaatcttaca ggtttccttc
     3961 aaaaccaaca agtcccacaa ataagaattc tgattaattc cgcccccccc cctcccccgc
     4021 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     4081 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     4141 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     4201 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     4261 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     4321 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     4381 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     4441 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     4501 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     4561 aaaaacacga tgataagctt gatcacgcgt ctcgagatga tccaccggag cttaccatga
     4621 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4681 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4741 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4801 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4861 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4921 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4981 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5041 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5101 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5161 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc
     5221 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgatcagtg
     5281 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     5341 tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc
     5401 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa
     5461 ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc gaccctagag
     5521 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     5581 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     5641 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
     5701 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     5761 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
     5821 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
     5881 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5941 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6001 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtag
     6061 atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     6121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     6181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     6241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tactggctta
     6421 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6481 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     6541 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6601 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6661 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6721 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6781 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6841 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6901 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6961 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7021 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7081 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     7141 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7201 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7261 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7321 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7381 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7441 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7501 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7561 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7621 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7681 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7741 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     7801 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7861 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7921 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7981 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8041 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     8101 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8161 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8221 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8281 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8341 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8401 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     8461 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     8521 aagaattagc ttggccattg catacgttgt atccatatca taatatgtac atttatattg
     8581 gctcatgtcc aacattaccg ccatgtt
//
