LOCUS       sequence-213694-        7018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gamma-Retroviral transfer vector for cloning and expressing your 
            gene of interest. IRES-driven Puromycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   Adamoski D, Dias MM, Quesnay JEN, Yang Z, Zagoriy I, Steyer AM, 
            Rodrigues CT, da Silva Bastos AC, da Silva BN, Costa RKE, de Abreu 
            FMO, Islam Z, Cassago A, van Heel MG, Consonni SR, Mattei S, Mahamid
            J, Portugal RV, Ambrosio ALB, Dias SMG
  TITLE     Molecular mechanism of glutaminase activation through filamentation 
            and the role of filaments in mitophagy protection.
  JOURNAL   Nat Struct Mol Biol. 2023 Dec;30(12):1902-1912. doi: 
            10.1038/s41594-023-01118-0. Epub 2023 Oct 19.
  PUBMED    37857822
REFERENCE   2  (bases 1 to 7018)
  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: 
            "10.1038/s41594-023-01118-0"; journalName: "Nat Struct Mol Biol"; 
            date: "2023-12"; volume: "30"; issue: "12"; pages: "1902-1912"
COMMENT     Sequence Label: pQC MCS IRES Puro
FEATURES             Location/Qualifiers
     source          1..7018
                     /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"
     misc_feature    2414..2987
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2581..2598)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2808..2827
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3028..3627
                     /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(3028..3047)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3524..3544
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3796..4221
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4324..4346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4482..4502)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4487..4816
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4667..4802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4838..4857
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4938..4955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5109..5697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5189..5208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5868..6659,6660..6728))
                     /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     6491..6510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6729..6833)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6901..6919
                     /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 ccggttctag
     2341 agccaccggt tgtatggagt ttaaactcca aagcatcgat ggtagcggcc gcggtgaatt
     2401 ctgattaatt ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     2461 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2521 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2581 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2641 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2701 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2761 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2821 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2881 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     2941 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct tgatcacgcg
     3001 tctcgagatg atccaccgga gcttaccatg accgagtaca agcccacggt gcgcctcgcc
     3061 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3121 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     3181 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     3241 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     3301 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     3361 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     3421 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     3481 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     3541 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     3601 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
     3661 aggagcgcac gaccccatgc atcgatcagt ggtccaggct ctagttttga ctcaacaata
     3721 tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct
     3781 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     3841 gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc
     3901 aggaacagat ggaacagggt cgaccctaga gaaccatcag atgtttccag ggtgccccaa
     3961 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     4021 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc
     4081 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     4141 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     4201 cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga
     4261 ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg atgacggtga
     4321 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4381 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     4441 gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt gtcagttagg
     4501 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4561 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4621 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4681 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4741 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4801 cctaggcttt tgcaaaaagc ttactggctt aactatgcgg catcagagca gattgtactg
     4861 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4921 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4981 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5041 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5101 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5161 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5221 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5281 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5341 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5401 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5461 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5521 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5581 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5641 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5701 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5761 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5821 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5881 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5941 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6001 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6061 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6121 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6181 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6241 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6301 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6361 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6421 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6481 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6541 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6601 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6661 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6721 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6781 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6841 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     6901 aataggcgta tcacgaggcc ctttcgtctt caagaattag cttggccatt gcatacgttg
     6961 tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc gccatgtt
//
