LOCUS       sequence-260904-        7157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SHMT2 K280A catalytic site mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7157)
  AUTHORS   Minton DR, Nam M, McLaughlin DJ, Shin J, Bayraktar EC, Alvarez SW, 
            Sviderskiy VO, Papagiannakopoulos T, Sabatini DM, Birsoy K, 
            Possemato R
  TITLE     Serine Catabolism by SHMT2 Is Required for Proper Mitochondrial 
            Translation Initiation and Maintenance of Formylmethionyl-tRNAs.
  JOURNAL   Mol Cell. 2018 Feb 15;69(4):610-621.e5. doi: 
            10.1016/j.molcel.2018.01.024.
  PUBMED    29452640
REFERENCE   2  (bases 1 to 7157)
  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.molcel.2018.01"; journalName: "Mol Cell"; date: 
            "2018-02-15- 15"; volume: "69"; issue: "4"; pages: "610-621.e5"
COMMENT     Sequence Label: pMXS-IRES-Blast SHMT2 CD
FEATURES             Location/Qualifiers
     source          1..7157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..498
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     428..450
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     466..482
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    508..882
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     820..836
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    2506..3070
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2520..3072
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2664..2681)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2891..2910
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3084..3479
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3650..4119
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4249..4266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4420..5008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4500..4519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5172..5963,5964..6032))
                     /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     5795..5814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6033..6137)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6167..6760
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    join(6823..7157,1..23)
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
       61 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
      121 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
      181 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
      241 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
      301 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
      361 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
      421 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
      481 actccttctc taggcgcccc catatggcca tatgagatct tatatggggc acccccgccc
      541 cttgtaaact tccctgaccc tgacatgaca agagttacta acagcccctc tctccaagct
      601 cacttacagg ctctctactt agtccagcac gaagtctgga gacctctggc ggcagcctac
      661 caagaacaac tggaccgacc ggtggtacct cacccttacc gagtcggcga cacagtgtgg
      721 gtccgccgac accagactaa gaacctagaa cctcgctgga aaggacctta cacagtcctg
      781 ctgaccaccc ccaccgccct caaagtagac ggcatcgcag cttggataca cgccgcccac
      841 gtgaaggctg ccgaccccgg gggtggacca tcctctagac tgccggatct agctagttaa
      901 ttaaggatcc cagtgtggtg gtacgggaat tcaccatgct gtacttctct ttgttttggg
      961 cggctcggcc tctgcagaga tgtgggcagc tggtcaggat ggccattcgg gctcagcaca
     1021 gcaacgcagc ccagactcag actggggaag caaacagggg ctggacaggc caggagagcc
     1081 tgtcggacag tgatcctgag atgtgggagt tgctgcagag ggagaaggac aggcaatgca
     1141 gaggcctgga gctcattgcc tcagagaact tctgcagccg agctgcgctg gaggccctgg
     1201 ggtcctgtct gaacaacaag tactcggagg gttatcctgg caagagatac tatgggggag
     1261 cagaggtggt ggatgaaatt gagctgctgt gccagcgccg ggccttggaa gcctttgacc
     1321 tggatcctgc acagtgggga gtcaatgtcc agccctactc cgggtcgcca gccaacctgg
     1381 ccgtctacac agcccttctg caacctcacg accggatcat ggggctggac ctgcccgatg
     1441 ggggccatct cacccacggc tacatgtctg acgtcaagcg gatatcagcc acgtccatct
     1501 tcttcgagtc tatgccctat aagctcaacc ccaaaactgg cctcattgac tacaaccagc
     1561 tggcactgac tgctcgactt ttccggccac ggctcatcat agctggcacc agcgcctatg
     1621 ctcgcctcat tgactacgcc cgcatgagag aggtgtgtga tgaagtcaaa gcacacctgc
     1681 tggcagacat ggcccacatc agtggcctgg tggctgccaa ggtgattccc tcgcctttca
     1741 agcacgcgga catcgtcacc accactactc acgcaactct tcgaggggcc aggtcagggc
     1801 tcatcttcta ccggaaaggg gtgaaggctg tggaccccaa gactggccgg gagatccctt
     1861 acacatttga ggaccgaatc aactttgccg tgttcccatc cctgcagggg ggcccccaca
     1921 atcatgccat tgctgcagta gctgtggccc taaagcaggc ctgcaccccc atgttccggg
     1981 agtactccct gcaggttctg aagaatgctc gggccatggc agatgccctg ctagagcgag
     2041 gctactcact ggtatcaggt ggtactgaca accacctggt gctggtggac ctgcggccca
     2101 agggcctgga tggagctcgg gctgagcggg tgctagagct tgtatccatc actgccaaca
     2161 agaacacctg tcctggagac cgaagtgcca tcacaccggg cggcctgcgg cttggggccc
     2221 cagccttaac ttctcgacag ttccgtgagg atgacttccg gagagttgtg gactttatag
     2281 atgaaggggt caacattggc ttagaggtga agagcaagac tgccaagctc caggatttca
     2341 aatccttcct gcttaaggac tcagaaacaa gtcagcgtct ggccaacctc aggcaacggg
     2401 tggagcagtt tgccagggcc ttccccatgc ctggttttga tgagcattga ctcgagggcc
     2461 ggcgcgccgc ggccgctacg taaattccgc cccccccccc cytcntccnc ccccccctaa
     2521 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     2581 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     2641 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     2701 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     2761 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     2821 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     2881 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     2941 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     3001 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     3061 acgatgataa tatggccaca accatggttc ctttgtctca agaagaatcc accctcattg
     3121 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     3181 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     3241 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     3301 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     3361 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     3421 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag
     3481 tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     3541 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     3601 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
     3661 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
     3721 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     3781 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
     3841 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     3901 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     3961 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     4021 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     4081 cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt
     4141 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     4201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5101 cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact
     5161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6001 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcataatg aaagacccca
     6181 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     6241 aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     6301 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     6361 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     6421 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     6481 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     6541 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     6601 aacccctcac tcggcgcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     6661 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     6721 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     6781 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
     6841 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
     6901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tcggaacacc
     6961 cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg gcccgacctg
     7021 agtccaaaaa tcccgatcgt tttggactct ttggtgcacc cccctaatag gagggatatg
     7081 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     7141 cggtttggga ccgaagc
//
