LOCUS       sequence-231460-        8541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses gpASNase1 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8541)
  AUTHORS   Sullivan LB, Luengo A, Danai LV, Bush LN, Diehl FF, Hosios AM, Lau 
            AN, Elmiligy S, Malstrom S, Lewis CA, Vander Heiden MG
  TITLE     Aspartate is an endogenous metabolic limitation for tumour growth.
  JOURNAL   Nat Cell Biol. 2018 Jul;20(7):782-788. doi: 
            10.1038/s41556-018-0125-0. Epub 2018 Jun 25.
  PUBMED    29941931
REFERENCE   2  (bases 1 to 8541)
  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/s41556-018-0125-0"; journalName: "Nat Cell Biol"; date: 
            "2018-07"; volume: "20"; issue: "7"; pages: "782-788"
COMMENT     Sequence Label: pLHCX-gpASNase1
FEATURES             Location/Qualifiers
     source          1..8541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             414..589
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    652..851
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1052..1468
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1398..1420
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1436..1452
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1524..2543
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             1524..2543
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     enhancer        2981..3360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3361..3564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3515..3535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3561..3585
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5371..5394
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             5443..6036
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6139..6161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6247..6387
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6302..6321
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6402..6419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6573..7161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6653..6672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7332..8123,8124..8192))
                     /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     7955..7974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8193..8297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8365..8383
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
     1441 tccagccctc actccttctc taggcgccgg aattccgatc tgatagcttg ccacaacccg
     1501 taccaaagat ggatagatcc ggaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     1561 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     1621 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     1681 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     1741 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     1801 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     1861 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     1921 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     1981 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     2041 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     2101 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     2161 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     2221 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     2281 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     2341 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     2401 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     2461 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     2521 ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa caagagctga tttcgagaac
     2581 gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg gccttacgct
     2641 tggtggcaca gttctcgtcc acagttcgct aagctcgctc ggctgggtcg cgggagggcc
     2701 ggtcgcagtg attcaggccc ttctggattg tgttggtccc cagggcacga ttgtcatgcc
     2761 cacgcactcg ggtgatctga ctgatcccgc agattggaga tcgccgcccg tgcctgccga
     2821 ttgggtgcag atctctagag ctcgctgatc cggccattag ccatattatt cattggttat
     2881 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatccata tcataatatg
     2941 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
     3001 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     3061 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     3121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     3181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     3241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     3301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     3361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     3421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     3481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     3541 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     3601 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccccaag
     3661 cttatggccc gagcgagtgg tagcgaaagg catttgctcc ttatttatac aggggggacg
     3721 ctggggatgc agtcaaaggg gggggtcctt gtacctgggc caggtctcgt gactctcctt
     3781 cgcacattgc cgatgttcca cgataaagag tttgcgcagg ctcaaggact gccggaccat
     3841 gctctcgcat tgcctcctgc ctcacacggt ccgcgggtcc tttacactgt attggagtgc
     3901 cagccacttc tggacagtag cgacatgaca atcgacgact ggattcggat tgccaagatt
     3961 attgaacgcc attacgaaca atatcagggt tttgttgtta tccatggtac agatacaatg
     4021 gcctcagggg caagtatgct gtcattcatg cttgaaaacc ttcataagcc ggtgatactc
     4081 acaggagctc aggtaccaat acgggtgttg tggaacgatg ccagggaaaa cctgcttggt
     4141 gcgcttctcg tggccggtca gtatatcatc cccgaggttt gcctttttat gaacagtcag
     4201 ctttttaggg gtaatcgagt gacaaaagtg gacagtcaga aatttgaagc cttctgcagc
     4261 cctaacctga gcccattggc tactgtgggg gctgatgtga ctatcgcctg ggacctcgtg
     4321 cgcaaagtaa agtggaagga cccgctcgtc gttcactcta atatggaaca tgacgtcgct
     4381 ctcttgaggc tgtatccagg gattccggca agtttggtac gcgcattcct gcaacctcct
     4441 ctcaaaggcg ttgtactcga gacttttgga tctggaaacg gcccgagcaa accggatctg
     4501 ttgcaagaac ttcgggcggc tgcacagcgg gggttgataa tggtaaattg ttctcagtgc
     4561 ttgagaggtt cagtgactcc tggttatgct acttccctcg ctggtgcaaa tatcgtgagt
     4621 ggcctcgaca tgactagtga ggctgccctt gctaagttgt cttacgtcct tggacttcca
     4681 gagttgtcat tggaaagaag gcaggagctc ctggctaagg acctgagagg tgagatgacc
     4741 ctccccactg ccgacctcca tcaaagttct ccacctggct ctacattggg tcaaggagtt
     4801 gcacgactgt tttcactttt tggatgccag gaagaggaca gtgtccagga tgcagttatg
     4861 ccttctttgg cattggcgct cgcgcatgca ggagagcttg aggcactcca ggcgctcatg
     4921 gaattgggat cagatctccg actcaaggac tcaaacggtc agaccctgct ccacgtggcg
     4981 gcaagaaacg ggcgagacgg agtggtcacg atgcttctcc atcgcggtat ggatgtaaac
     5041 gcgagagatc gcgacggact gagtcccctc ctgttggcgg tccaggggag acatcgagag
     5101 tgtatacgac tgcttcgcaa agctggggct tgcttgtctc cacaagacct caaagatgca
     5161 ggaactgaac tttgtcggct ggcgagcaga gcagacatgg aggggctgca ggcttgggga
     5221 caggcgggag cggacctcca gcagcctggt tacgacggca ggagcgccct ttgcgtggct
     5281 gaagcggccg gaaatcaaga agtgctggcg ctccttagga acctggcact cgtgggacca
     5341 gaggtgccac ctgccattgg ttctggttct gactacaaag acgatgacga caagtagatc
     5401 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     5461 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     5521 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     5581 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     5641 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     5701 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     5761 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     5821 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     5881 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     5941 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     6001 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag
     6061 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt
     6121 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6181 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6241 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     6301 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     6361 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     6421 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6481 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6541 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6601 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6661 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6721 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6781 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6841 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6901 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6961 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7021 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7081 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7141 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7201 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7261 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7321 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7381 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7441 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7501 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7561 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7621 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     7681 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7741 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7801 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7861 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7921 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     7981 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8041 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8101 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8161 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8221 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8281 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8341 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa
     8401 ttcataccag atcaccgaaa actgtcctcc aaatgtgtcc ccctcacact cccaaattcg
     8461 cgggcttctg cctcttagac cactctaccc tattccccac actcaccgga gccaaagccg
     8521 cggcccttcc gtttctttgc t
//
