LOCUS       sequence-207713-        7423 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 7423)
  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 7423)
  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..7423
                     /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"
     misc_feature    join(2332..2905,2906..2908,2909..2917)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2499..2516)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2726..2745
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2918..3712
                     /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(2972..2991)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3582..3601
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4129..4554
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4657..4679)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4815..4835)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4820..5149
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5000..5135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5171..5190
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5271..5288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5442..6030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5522..5541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6201..6992,6993..7061))
                     /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     6824..6843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7062..7166)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7234..7252
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7352..7423,1..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 ccggggatcc ggttctagag ccaccggttg
     2281 tatggagttt aaactccaaa gcatcgatgg tagcggccgc ggtgaattcc gcccctctcc
     2341 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2401 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2461 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     2521 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     2581 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     2641 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     2701 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     2761 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     2821 tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     2881 ttcctttgaa aaacacgatg ataagcttgc cacaaccatg gctgaacaag atggattgca
     2941 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3001 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3061 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3121 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3181 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3241 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3301 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3361 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3421 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3481 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3541 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3601 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3661 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggatc
     3721 ggctagagtc gacccggacg ggacgctcct gcgcctgata cagaacgaat tgcttgcagg
     3781 catctcatga gtgtgtcttc ccgttttccg cctgaggtca ctgcgtggat ggagcgctgg
     3841 cgcctgctgc gcgacggcga gctgctcacc acccactcga gggctgcagc gctgcagagg
     3901 ccgagtgcag aactgctcca aagggacctc aaggctttcc gagggacact aggctgactc
     3961 catcgagcca gtgtagagat aagcttatcg attagtccaa tttgttaaag acaggatatc
     4021 agtggtccag gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga
     4081 gccatagata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     4141 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     4201 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ggtcgaccct
     4261 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     4321 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     4381 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     4441 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     4501 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
     4561 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
     4621 tggctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4681 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4741 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4801 gtagatccgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4861 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4921 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4981 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5041 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5101 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttactgg
     5161 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5221 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     5281 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5341 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5401 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5461 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5521 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5581 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5641 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5701 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5761 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5821 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5881 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5941 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6001 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6061 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6121 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6181 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6241 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6301 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6361 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6421 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6481 ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg
     6541 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6601 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6661 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6721 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6781 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     6841 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6901 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6961 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7021 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7081 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7141 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7201 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7261 cttcaagaat tagcttggcc attgcatacg ttgtatccat atcataatat gtacatttat
     7321 attggctcat gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag
     7381 taatcaatta cggggtcatt agttcatagc ccatatatgg agt
//
