LOCUS       sequence-379337-        8414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector used to immortalize human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8414)
  AUTHORS   Shi DD, Savani MR, Levitt MM, Wang AC, Endress JE, Bird CE, Buehler 
            J, Stopka SA, Regan MS, Lin YF, Puliyappadamba VT, Gao W, Khanal J, 
            Evans L, Lee JH, Guo L, Xiao Y, Xu M, Huang B, Jennings RB, Bonal 
            DM, Martin-Sandoval MS, Dang T, Gattie LC, Cameron AB, Lee S, Asara 
            JM, Kornblum HI, Mak TW, Looper RE, Nguyen QD, Signoretti S, Gradl 
            S, Sutter A, Jeffers M, Janzer A, Lehrman MA, Zacharias LG, Mathews 
            TP, Losman JA, Richardson TE, Cahill DP, DeBerardinis RJ, Ligon KL, 
            Xu L, Ly P, Agar NYR, Abdullah KG, Harris IS, Kaelin WG Jr, McBrayer
            SK
  TITLE     De novo pyrimidine synthesis is a targetable vulnerability in IDH 
            mutant glioma.
  JOURNAL   Cancer Cell. 2022 Sep 12;40(9):939-956.e16. doi: 
            10.1016/j.ccell.2022.07.011. Epub 2022 Aug 18.
  PUBMED    35985343
REFERENCE   2  (bases 1 to 8414)
  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: 
            "10.1016/j.ccell.2022.07.011"; journalName: "Cancer Cell"; date: 
            "2022-09-12- 12"; volume: "40"; issue: "9"; pages: "939-956.e16"
COMMENT     Sequence Label: pLBCX-Large-T-antigenK1 mutant
FEATURES             Location/Qualifiers
     source          1..8414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             217..392
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    455..654
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             855..1271
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1201..1223
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1239..1255
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1314..1712
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     enhancer        2194..2573
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2574..2777
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2728..2748
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2774..2798
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2912..5038
                     /codon_start=1
                     /product="large T antigen from SV40 (simian virus 40)"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSKEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     LTR             5119..5712
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5815..5837)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5923..6063
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5978..5997
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6078..6095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6249..6837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6329..6348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7008..7799,7800..7868))
                     /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     7631..7650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7869..7973)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8041..8059
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcgggg ccaagaacag
       61 atggtcccca gatgcggtcc agccctcagc agtttctagt gaatcatcag atgtttccag
      121 ggtgccccaa ggacctgaaa atgaccctgt accttatttg aactaaccaa tcagttcgct
      181 tctcgcttct gttcgcgcgc ttccgctctc cgagctcaat aaaagagccc acaacccctc
      241 actcggcgcg ccagtcttcc gatagactgc gtcgcccggg tacccgtatt cccaataaag
      301 cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc ttgggagggt ctcctctgag
      361 tgattgacta cccacgacgg gggtctttca tttgggggct cgtccgggat ttggagaccc
      421 ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact tatctgtgtc
      481 tgtccgattg tctagtgtct atgtttgatg ttatgcgcct gcgtctgtac tagttagcta
      541 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
      601 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
      661 agtcgatgtg gaatccgacc ccgtcaggat atgtggttct ggtaggagac gagaacctaa
      721 aacagttccc gcctccgtct gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt
      781 gtctgctgca gcgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
      841 tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta ggtcactgga
      901 aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga cgttgggtta
      961 ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac ggcaccttta
     1021 accgagacct catcacccag gttaagatca aggtcttttc acctggcccg catggacacc
     1081 cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc cctccctggg
     1141 tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc
     1201 cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc ctcactcctt
     1261 ctctaggcgc cggaattccg atctgatcaa gagacaggat ggggtcgttt cgcatggcca
     1321 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     1381 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     1441 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     1501 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     1561 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     1621 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     1681 tgctgccctc tggttatgtg tgggagggct aagcgggact ctggggttcg aaatgaccga
     1741 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     1801 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     1861 catgctggag ttcttcgccc accccgggct cgatcccctc gcgagttggt tcagctgctg
     1921 cctgaggctg gacgacctcg cggagttcta ccggcagtgc aaatccgtcg gcatccagga
     1981 aaccagcagc ggctatccgc gcatccatgc ccccgaactg caggagtggg gaggcacgat
     2041 ggccgctttg gtcgaggcgg atccggccat tagccatatt attcattggt tatatagcat
     2101 aaatcaatat tggctattgg ccattgcata cgttgtatcc atatcataat atgtacattt
     2161 atattggctc atgtccaaca ttaccgccat gttgacattg attattgact agttattaat
     2221 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2281 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2341 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2401 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2461 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2521 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2581 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2641 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     2701 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     2761 tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct
     2821 gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggcccc aagcttgttg
     2881 atcccgcgac cggtcgacca gctttgcaaa gatggataaa gttttaaaca gagaggaatc
     2941 tttgcagcta atggaccttc taggtcttga aaggagtgcc tgggggaata ttcctctgat
     3001 gagaaaggca tatttaaaaa aatgcaagga gtttcatcct gataaaggag gagatgaaga
     3061 aaaaatgaag aaaatgaata ctctgtacaa gaaaatggaa gatggagtaa aatatgctca
     3121 tcaacctgac tttggaggct tctgggatgc aactgagatt ccaacctatg gaactgatga
     3181 atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcaa aagaaatgcc
     3241 atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag
     3301 aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt
     3361 gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact
     3421 gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta
     3481 taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa
     3541 ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata
     3601 tttgatgtat agtgccttga ctagagatcc attttctgtt attgaggaaa gtttgccagg
     3661 tgggttaaag gagcatgatt ttaatccaga agaagcagag gaaactaaac aagtgtcctg
     3721 gaagcttgta acagagtatg caatggaaac aaaatgtgat gatgtgttgt tattgcttgg
     3781 gatgtacttg gaatttcagt acagttttga aatgtgttta aaatgtatta aaaaagaaca
     3841 gcccagccac tataagtacc atgaaaagca ttatgcaaat gctgctatat ttgctgacag
     3901 caaaaaccaa aaaaccatat gccaacaggc tgttgatact gttttagcta aaaagcgggt
     3961 tgatagccta caattaacta gagaacaaat gttaacaaac agatttaatg atcttttgga
     4021 taggatggat ataatgtttg gttctacagg ctctgctgac atagaagaat ggatggctgg
     4081 agttgcttgg ctacactgtt tgttgcccaa aatggattca gtggtgtatg actttttaaa
     4141 atgcatggtg tacaacattc ctaaaaaaag atactggctg tttaaaggac caattgatag
     4201 tggtaaaact acattagcag ctgctttgct tgaattatgt ggggggaaag ctttaaatgt
     4261 taatttgccc ttggacaggc tgaactttga gctaggagta gctattgacc agtttttagt
     4321 agtttttgag gatgtaaagg gcactggagg ggagtccaga gatttgcctt caggtcaggg
     4381 aattaataac ctggacaatt taagggatta tttggatggc agtgttaagg taaacttaga
     4441 aaagaaacac ctaaataaaa gaactcaaat atttccccct ggaatagtca ccatgaatga
     4501 gtacagtgtg cctaaaacac tgcaggccag atttgtaaaa caaatagatt ttaggcccaa
     4561 agattattta aagcattgcc tggaacgcag tgagtttttg ttagaaaaga gaataattca
     4621 aagtggcatt gctttgcttc ttatgttaat ttggtacaga cctgtggctg agtttgctca
     4681 aagtattcag agcagaattg tggagtggaa agagagattg gacaaagagt ttagtttgtc
     4741 agtgtatcaa aaaatgaagt ttaatgtggc tatgggaatt ggagttttag attggctaag
     4801 aaacagtgat gatgatgatg aagacagcca ggaaaatgct gataaaaatg aagatggtgg
     4861 ggagaagaac atggaagact cagggcatga aacaggcatt gattcacagt cccaaggctc
     4921 atttcaggcc cctcagtcct cacagtctgt tcatgatcat aatcagccat accacatttg
     4981 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     5041 tgaatgcaat tgttgttgtt aacggggatc aacatcgata aaataaaaga ttttatttag
     5101 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     5161 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     5221 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     5281 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     5341 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     5401 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     5461 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     5521 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     5581 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     5641 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     5701 gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc
     5761 caccaccggg aggtaagctg gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     5821 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5881 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     5941 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     6001 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     6061 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6121 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6181 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6241 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6301 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6361 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6421 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6481 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6541 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6601 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6661 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     6721 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6781 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6841 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6961 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7021 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7081 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7141 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7201 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7261 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7321 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7381 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7441 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7501 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7561 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7621 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7681 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7741 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7801 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7861 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7921 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7981 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     8041 aataggcgta tcacgaggcc ctttcgtctt caagaattca taccagatca ccgaaaactg
     8101 tcctccaaat gtgtccccct cacactccca aattcgcggg cttctgcctc ttagaccact
     8161 ctaccctatt ccccacactc accggagcca aagccgcggc ccttccgttt ctttgctttt
     8221 gaaagacccc acccgtaggt ggcaagctag cttaagtaac gccactttgc aaggcatgga
     8281 aaaatacata actgagaata gaaaagttca gatcaaggtc aggaacaaag aaacagctga
     8341 ataccaaaca ggatatctgt ggtaagcggt tcctgccccg gctcagggcc aagaacagat
     8401 gagacagctg agtg
//
