LOCUS       sequence-213718-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gamma-Retroviral transfer vector for cloning and expressing your 
            gene of interest. IRES-driven Geneticin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  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 7424)
  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 G418
FEATURES             Location/Qualifiers
     source          1..7424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             586..761
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    824..1023
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1224..1640
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1570..1592
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1608..1624
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1677..1980
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1981..2184
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2181..2205
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    join(2405..2979,2980..2982,2983..2991)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2572..2589)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2799..2818
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2992..3786
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3046..3065)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3656..3675
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4203..4628
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4731..4753)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4889..4909)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4894..5223
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5074..5209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5245..5264
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5345..5362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5516..6104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5596..5615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6275..7066,7067..7135))
                     /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     6898..6917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7136..7240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7308..7326
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata aaagagccca
      601 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
      661 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
      721 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
      781 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
      841 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
      901 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
      961 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     1021 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     1081 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     1141 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     1201 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     1261 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     1321 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     1381 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     1441 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     1501 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     1561 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     1621 tcactccttc tctaggcgcc ggaattgaag atctggggga tcgatcctct agagtccgtt
     1681 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1741 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1801 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1861 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1921 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1981 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2041 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2101 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2161 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2221 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccgggaa
     2281 cggtgcattg gaacggacct gcaggaattg atccgcggcc gcaccgggga tccggttcta
     2341 gagccaccgg ttgtatggag tttaaactcc aaagcatcga tggtagcggc cgcggtgaat
     2401 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     2461 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     2521 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     2581 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     2641 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     2701 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     2761 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     2821 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     2881 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     2941 acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac catggctgaa
     3001 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3061 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3121 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3181 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3241 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3301 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3361 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3421 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3481 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3541 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3601 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3661 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3721 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3781 ttctgagcgg gatcggctag agtcgacccg gacgggacgc tcctgcgcct gatacagaac
     3841 gaattgcttg caggcatctc atgagtgtgt cttcccgttt tccgcctgag gtcactgcgt
     3901 ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct caccacccac tcgagggctg
     3961 cagcgctgca gaggccgagt gcagaactgc tccaaaggga cctcaaggct ttccgaggga
     4021 cactaggctg actccatcga gccagtgtag agataagctt atcgattagt ccaatttgtt
     4081 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     4141 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     4201 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4261 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     4321 acagggtcga ccctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     4381 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     4441 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     4501 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     4561 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     4621 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
     4681 accgggaggt aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4741 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4801 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     4861 agcgatagcg gagtgtagat ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4921 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5101 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5161 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5221 aaaaagctta ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5281 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     5341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5941 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     6601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     6901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7201 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7261 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     7321 cgaggccctt tcgtcttcaa gaattagctt ggccattgca tacgttgtat ccatatcata
     7381 atatgtacat ttatattggc tcatgtccaa cattaccgcc atgt
//
