LOCUS       sequence-380517-        7906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Asparagine synthetase overexpression in murine cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  AUTHORS   Fernandez-Garcia J, Franco F, Parik S, Altea-Manzano P, Pane AA, 
            Broekaert D, van Elsen J, Vermeire I, Schalley T, Planque M, van 
            Brussel T, Schepers R, Modave E, Karakach TK, Carmeliet P, 
            Lambrechts D, Ho PC, Fendt SM
  TITLE     CD8(+) T cell metabolic rewiring defined by scRNA-seq identifies a 
            critical role of ASNS expression dynamics in T cell differentiation.
  JOURNAL   Cell Rep. 2022 Nov 15;41(7):111639. doi: 
            10.1016/j.celrep.2022.111639.
  PUBMED    36384124
REFERENCE   2  (bases 1 to 7906)
  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"; journalName: "Cell Rep.";
            date: "2022-11-15"; volume: "41(7):111639. doi"; pages: " 
            10.1016/j.celrep.2022.111639"
COMMENT     Sequence Label: MSCV-ASNS-OE-IRES-Thy1.1
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     211..233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     361..380
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             580..1096
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1160..1501
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1568..1984
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1914..1936
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1952..1968
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    3720..4286
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3736..4288
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3880..3897)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4107..4126
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4336..4824
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     LTR             4880..5394
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5556..5572
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5580..5586,5587..5604,5605..5610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5625..5646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5763..5780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5934..6522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6014..6033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6693..7484,7485..7553))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7316..7335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7554..7658)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7726..7744
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7782..7804)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(7904..7906,1..17)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
       61 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
      121 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      181 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc
      241 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
      301 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
      361 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
      421 gatgcgtccg gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca
      481 ggctctagtt ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat
      541 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
      601 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
      661 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
      721 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
      781 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      841 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      901 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      961 ctccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca
     1021 tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc
     1081 tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc
     1141 gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgt
     1201 gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag ctaactagct
     1261 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
     1321 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
     1381 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
     1441 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
     1501 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     1561 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     1621 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     1681 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     1741 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     1801 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     1861 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     1921 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     1981 cgccggaatt agatctgcca ccatgtgtgg catctgggcc ctcttcggca gcgatgactg
     2041 cctttccgtg cagtgtctga gtgcgatgaa gatcgcgcac agggggccag atgcatttcg
     2101 ctttgagaat gtcaatggat acaccaactg ctgctttggc tttcaccgct tggctgtggt
     2161 tgacccgctg tttggaatgc agccgataag agtgaggaaa tacccttatt tgtggctctg
     2221 ttacaatggt gaaatctaca accacaaggc gctacagcaa cgttttgaat ttgaatatca
     2281 gaccaatgtg gatggtgaga ttatcctcca cctctatgac aaaggaggca tcgagaaaac
     2341 catctgtatg ctggacgggg tgtttgcatt catcttactg gacactgcca ataagaaagt
     2401 atttctgggc agagacacct atggagtcag gcccttgttt aaagccatga cagaagatgg
     2461 gtttctggct gtgtgttcag aagctaaagg ccttgtttcc ttaaaacact ccaccactcc
     2521 cttcttaaaa gtggagccct tccttcctgg acactatgaa gttttggatt taaaaccaaa
     2581 tggcaaagtt gcgtctgtgg aaatggtcaa ataccatcac tgtacggatg aaccattgca
     2641 tgccatctat gacagcgtgg agaaactctt cccaggcttt gacctagaga ccgtgaagaa
     2701 caatctgcgt atcctttttg acaacgctat caagaaacgc ttgatgacag accggaggat
     2761 tggctgcctt ttatcagggg gcctggactc gagcttggtt gctgcctctc tgctgaagca
     2821 actcaaggag gcccaagttc agtatcctct ccagacattt gctattggca tggaggacag
     2881 ccccgatctc ctggccgcta gaaaggtggc aaattatatt ggaagcgagc atcatgaagt
     2941 cctttttaac tctgaagaag gcattcaggc cctggatgaa gtcatatttt ccttggaaac
     3001 ttatgatatt acgacagttc gggcatctgt gggcatgtat ttaatttcca agtatattcg
     3061 gaagaacaca gacagcgtgg tgatcttctc cggagagggg tcagatgaac ttacacaggg
     3121 ctatatatat ttccacaagg ctccttcccc tgagaaggcc gaggaggaga gtgagagact
     3181 gctgaaggaa ctctacctgt ttgatgttct ccgggccgac cgcactactg ctgcacacgg
     3241 tctcgaactg agagtcccct ttctggatca tcggttttct tcctattacc tgtctctgcc
     3301 gccagatatg agaattccaa aaaatggcat agaaaaacat ctcctgagag agacttttga
     3361 ggactgcaac ctgctaccca aagagattct ctggcgaccc aaagaagcct tcagtgatgg
     3421 gatcacctca gtcaagaact cctggttcaa gattttgcag gactatgttg aacatcaggt
     3481 tgatgatgag atgatgtctg cagcctccca gaagtttccc ttcaatactc ccaaaactaa
     3541 ggaaggctac ttctaccgtc agatctttga acgccattac ccaggccggg ctgattggct
     3601 gactcattat tggatgccta agtggatcaa tgctactgac ccttctgccc gcactctgac
     3661 ccattataag tcagctgcca aagcttaggt cgacggtatc gataagcttg atatcgaatt
     3721 ccgccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     3781 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     3841 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3901 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     3961 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4021 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     4081 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     4141 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     4201 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     4261 ttttcctttg aaaaacacga tgataatatg gccacaacca tgggtctttt ctgcagtcac
     4321 cgtcctcgag gcaccatgaa cccagccatc agcgtcgctc tcctgctctc agtcttgcag
     4381 gtgtcccgag ggcagaaggt gaccagcctg acagcctgcc tggtgaacca aaaccttcgc
     4441 ctggactgcc gccatgagaa taacaccaag gataactcca tccagcatga gttcagcctg
     4501 acccgagaga agaggaagca cgtgctctca ggcacccttg ggatacccga gcacacgtac
     4561 cgctcccgcg tcaccctctc caaccagccc tatatcaagg tccttaccct agccaacttc
     4621 accaccaagg atgagggcga ctacttttgt gagcttcgcg tctcgggcgc gaatcccatg
     4681 agctccaata aaagtatcag tgtgtataga gacaagctgg tcaagtgtgg cggcataagc
     4741 ctgctggttc agaacacatc ctggatgctg ctgctgctgc tttccctctc cctcctccaa
     4801 gccctggact tcatttctct gtgacgcgta gaattcgata aaataaaaga ttttatttag
     4861 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     4921 aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt cagatcaagg
     4981 ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta agcagttcct
     5041 gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct cagcagtttc
     5101 tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat
     5161 ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc
     5221 aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc
     5281 gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt
     5341 ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa
     5401 cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct
     5461 gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat
     5521 ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac aattccacac
     5581 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5641 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5701 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5761 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5821 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7681 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7741 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     7801 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     7861 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcgg
//
