LOCUS       sequence-213721-        8006 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 8006)
  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  JOURNAL   Unpublished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 8006)
  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            "Unpublished"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..8006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1604..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2108..2132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2266..2307
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3330..3903
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3497..3514)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3724..3743
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3944..4543
                     /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(3944..3963)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4440..4460
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4712..5137
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5240..5262)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5398..5418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5403..5732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5583..5718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5754..5773
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5854..5871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6025..6613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6105..6124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6784..7575,7576..7644))
                     /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     7407..7426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7645..7749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7817..7835
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7935..8006,1..308)
                     /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 gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgggggatcg atcctctaga gtccgttaca taacttacgg
     1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1921 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2101 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2161 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
     2221 cggacctgca ggaattgatc cgcggccgca ccggggatca ccatgggtaa gcctatccct
     2281 aaccctctcc tcggtctcga ttctacggga tccggttcta gagccaccgg tatgtctaat
     2341 ggttatgaag accacatggc cgaagactgc aggggtgaca tcgggagaac gaatttgatc
     2401 gtcaactacc tccctcagaa catgacccag gatgagttac gaagcctgtt cagcagcatt
     2461 ggtgaagttg aatctgcaaa acttattcgg gataaagtag caggacacag cttgggctat
     2521 ggctttgtga actacgtgac cgcgaaggat gcagagagag cgatcaacac gctgaacggc
     2581 ttgaggctcc agtcaaaaac cattaaggtg tcgtatgctc gcccgagctc agaggtgatc
     2641 aaagacgcca acttgtacat cagcgggctc ccgcggacca tgacccagaa ggacgtagaa
     2701 gacatgttct ctcggtttgg gcggatcatc aactcgcggg tcctcgtgga tcagactaca
     2761 ggtttgtcca gaggggttgc gtttatccgg tttgacaaac ggtcggaggc agaagaggca
     2821 attaccagtt tcaatggtca taaaccccca ggttcctctg agcccatcac agtgaagttt
     2881 gcagccaacc ccaaccagaa caaaaacgtg gcactcctct cgcagctgta ccactcgcca
     2941 gcgcgacggt tcggaggccc cgttcaccac caggcgcaga gattcaggtt ctcccccatg
     3001 ggcgtcgatc acatgagcgg gctctctggc gtcaacgtgc caggaaacgc ctcctccggc
     3061 tggtgcattt tcatctacaa cctggggcag gatgccgacg aggggatcct ctggcagatg
     3121 tttgggccgt ttggtgccgt caccaatgtg aaagtgatcc gcgacttcaa caccaacaag
     3181 tgcaaagggt ttggctttgt gaccatgaca aactatgaag aagccgcgat ggccatagcc
     3241 agcctgaacg gctaccgcct gggggacaaa atcttacagg tttccttcaa aaccaacaag
     3301 tcccacaaat agaattctga ttaattccgc ccctctccct cccccccccc taacgttact
     3361 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     3421 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     3481 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     3541 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     3601 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     3661 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     3721 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3781 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3841 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3901 taagcttgat cacgcgtctc gagatgatcc accggagctt accatgaccg agtacaagcc
     3961 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4021 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4081 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4141 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4201 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4261 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4321 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4381 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4441 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4501 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     4561 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg atcagtggtc caggctctag
     4621 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     4681 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     4741 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     4801 agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac catcagatgt
     4861 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     4921 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     4981 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     5041 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     5101 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     5161 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     5221 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5281 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5341 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc cggctgtgga
     5401 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5461 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5521 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5581 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5641 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5701 gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact atgcggcatc
     5761 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     5821 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5881 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5941 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6001 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6061 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6121 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6181 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6241 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6301 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6361 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6421 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6481 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6541 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6601 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6661 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6721 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6781 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6841 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6901 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6961 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7021 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7081 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7141 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7201 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7261 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7321 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7381 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     7441 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7501 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7561 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7621 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7681 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7741 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7801 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattagcttg
     7861 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     7921 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     7981 attagttcat agcccatata tggagt
//
