LOCUS       sequence-212925-        6655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Flag-SFXN1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  AUTHORS   Kory N, Wyant GA, Prakash G, Uit de Bos J, Bottanelli F, Pacold ME, 
            Chan SH, Lewis CA, Wang T, Keys HR, Guo YE, Sabatini DM
  TITLE     SFXN1 is a mitochondrial serine transporter required for one-carbon 
            metabolism.
  JOURNAL   Science. 2018 Nov 16;362(6416). pii: 362/6416/eaat9528. doi: 
            10.1126/science.aat9528.
  PUBMED    30442778
REFERENCE   2  (bases 1 to 6655)
  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: "Science."; 
            date: "2018-11-16"; volume: "362(6416). pii: 362/6416/eaat9528. 
            doi"; pages: " 10.1126/science.aat9528"
COMMENT     Sequence Label: pMXs_FLAG-SFXN1
FEATURES             Location/Qualifiers
     source          1..6655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             58..651
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    714..1071
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1130..1546
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1476..1498
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1514..1530
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1556..1930
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1868..1884
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1959..1968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1968..2033
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3066..3618
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3210..3227)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3437..3456
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3630..4025
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4196..4665
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4795..4812)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4966..5554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5046..5065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5718..6509,6510..6578))
                     /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     6341..6360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6579..6655,1..28))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgcataat
       61 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
      121 gaaaaataca taactgagaa tagaaaagtt cagatcaagg tcaggaacag atggaacagc
      181 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      241 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
      301 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
      361 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
      421 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
      481 aaagagccca caacccctca ctcggcgcgc cagtcctccg attgactgag tcgcccgggt
      541 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
      601 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
      661 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
      721 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt tatgcgcctg
      781 cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
      841 ttcggaacac ccggccgcaa ccctgggaga cgtcccaggg acttcggggg ccgtttttgt
      901 ggcccgacct gagtccaaaa atcccgatcg ttttggactc tttggtgcac ccccctaata
      961 ggagggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     1021 atttttgctt tcggtttggg accgaagccg cgccgcgcgt cttgtctgct gcagcatcgt
     1081 tctgtgttgt ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt
     1141 taccactccc ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa
     1201 ccagtcggta gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac
     1261 ctttaacgtc ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa
     1321 gatcaaggtc ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac
     1381 ctgggaagcc ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc
     1441 tccgcctcct cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc
     1501 gcctcgatcc tccctttatc cagccctcac tccttctcta ggcgccccca tatggccata
     1561 tgagatctta tatggggcac ccccgcccct tgtaaacttc cctgaccctg acatgacaag
     1621 agttactaac agcccctctc tccaagctca cttacaggct ctctacttag tccagcacga
     1681 agtctggaga cctctggcgg cagcctacca agaacaactg gaccgaccgg tggtacctca
     1741 cccttaccga gtcggcgaca cagtgtgggt ccgccgacac cagactaaga acctagaacc
     1801 tcgctggaaa ggaccttaca cagtcctgct gaccaccccc accgccctca aagtagacgg
     1861 catcgcagct tggatacacg ccgcccacgt gaaggctgcc gaccccgggg gtggaccatc
     1921 ctctagactg ccggatctag ctagttaatt aaggatccgc caccatggac tacaaggacc
     1981 acgacggtga ctacaaggac cacgacatcg actacaagga cgacgacgac aaggggcatg
     2041 gcaccggcag caccggcagc gaattcatga gcggtgaact gcctccaaac ataaatatta
     2101 aggaaccacg atgggatcaa tctacgttca tcggaagggc taaccatttc tttacagtga
     2161 cagacccgag gaatattttg ttgaccaacg agcaattgga gtctgcaagg aaaatcgtcc
     2221 atgattaccg acaaggtatt gtgccgccag gacttactga aaacgaactt tggagagcga
     2281 aatacattta tgactctgca ttccatccag acactggaga aaagatgatc ctgattggtc
     2341 gcatgtcagc tcaagtcccc atgaatatga ctataactgg ctgtatgatg acattttaca
     2401 gaacaacacc cgctgttctc ttttggcaat ggataaatca gagcttcaac gccgtagtaa
     2461 actacacaaa cagatccggc gacgctccac tgactgtcaa cgaactcgga actgcctacg
     2521 tgtccgccac gactggggcc gtcgccaccg ccctgggact taatgcactg acgaagcatg
     2581 ttagccctct cataggccgg ttcgttccgt tcgcggccgt agcagccgct aactgtatca
     2641 acatacctct gatgagacag agggaactga aggtcggaat cccagtgacc gatgagaatg
     2701 gcaatcgact gggcgagagt gcaaacgcag ctaaacaggc gataacgcaa gtggtagtta
     2761 gtaggatatt gatggcggct cccggaatgg cgataccccc cttcattatg aacaccctgg
     2821 agaaaaaggc gtttttgaaa cgcttcccat ggatgtctgc gccaatacag gtcgggctcg
     2881 taggtttctg tttggtcttt gctactcctc tgtgttgcgc actcttcccc caaaaaagca
     2941 gtatgtcagt tacaagcctc gaggctgagc tccaagctaa aattcaggag tctcaccctg
     3001 aattgcgacg agtttacttt aataagggac tttaagcggc cgctacgtaa attccgcccc
     3061 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3121 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3181 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3241 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3301 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3361 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3421 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3481 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     3541 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     3601 aaaaacacga tgataatatg gccacaacca tggttccttt gtctcaagaa gaatccaccc
     3661 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
     3721 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
     3781 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
     3841 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
     3901 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
     3961 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
     4021 gctaagtcga cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     4081 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     4141 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
     4201 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4261 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     4321 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     4381 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     4441 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     4501 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     4561 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     4621 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc
     4681 aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg
     4741 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4801 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4861 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4921 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4981 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5041 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5101 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5161 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5221 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5281 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5341 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5401 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5461 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5521 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5581 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5641 aaggatcttc acctagatcc ttttgcggcc ggccgcaaat caatctaaag tatatatgag
     5701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6301 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tattt
//
