LOCUS       sequence-214939-        6722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Flag-SFXN4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6722)
  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 6722)
  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-SFXN4
FEATURES             Location/Qualifiers
     source          1..6722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             21..614
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    677..1034
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1093..1509
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1439..1461
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1477..1493
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1519..1893
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1831..1847
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1922..1931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1931..1996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3074..3647,3648..3650,3651..3659)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3240..3257)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3467..3486
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3660..4055
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4226..4695
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4825..4842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4996..5584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5076..5095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5748..6539,6540..6608))
                     /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     6371..6390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6609..6713)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgcat aatgaaagac cccacctgta ggtttggcaa gctagcttaa
       61 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagaaaa gttcagatca
      121 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      181 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
      241 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
      301 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
      361 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
      421 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
      481 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
      541 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
      601 gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga
      661 cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt
      721 ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc
      781 ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg agacgtccca
      841 gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga tcgttttgga
      901 ctctttggtg caccccccta ataggaggga tatgtggttc tggtaggaga cgagaaccta
      961 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg
     1021 cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg
     1081 tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa
     1141 gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc
     1201 ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac
     1261 cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca
     1321 gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc
     1381 aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc
     1441 cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct
     1501 ctaggcgccc ccatatggcc atatgagatc ttatatgggg cacccccgcc ccttgtaaac
     1561 ttccctgacc ctgacatgac aagagttact aacagcccct ctctccaagc tcacttacag
     1621 gctctctact tagtccagca cgaagtctgg agacctctgg cggcagccta ccaagaacaa
     1681 ctggaccgac cggtggtacc tcacccttac cgagtcggcg acacagtgtg ggtccgccga
     1741 caccagacta agaacctaga acctcgctgg aaaggacctt acacagtcct gctgaccacc
     1801 cccaccgccc tcaaagtaga cggcatcgca gcttggatac acgccgccca cgtgaaggct
     1861 gccgaccccg ggggtggacc atcctctaga ctgccggatc tagctagtta attaaggatc
     1921 cgccaccatg gactacaagg accacgacgg tgactacaag gaccacgaca tcgactacaa
     1981 ggacgacgac gacaaggggc atggcaccgg cagcaccggc agcgaattca tgtccctgga
     2041 acaggaggag gaaacgcaac ctgggcggct cctaggacgc agagacgccg tccccgcctt
     2101 cattgagccc aacgtgcgct tctggatcac cgagcgccaa tcctttattc gacgatttct
     2161 tcaatggaca gaattattag atcctacaaa tgtgttcatt tcagttgaaa gtatagaaaa
     2221 ctcgaggcaa ctattgtgca caaatgaaga tgtttccagc cctgcctcgg cggaccaaag
     2281 gatacaggaa gcttggaagc ggagtcttgc aacagtgcat cccgacagca gcaacctgat
     2341 ccccaagctt tttcgacctg cagcgttcct gcctttcatg gcgcccacgg tatttttgtc
     2401 aatgacgcca ctgaaaggga tcaagtccgt gattttacct caggttttcc tctgtgccta
     2461 catggcagcg ttcaacagca tcaatggaaa cagaagttac acttgtaagc cactagaaag
     2521 atcattacta atggcgggag ccgttgcttc ttcaactttc ttaggagtaa tccctcagtt
     2581 tgtccagatg aagtatggcc tgactggccc ttggattaaa agactcttac ctgtgatctt
     2641 cctcgtgcaa gccagtggaa tgaatgtcta catgtcccga agtcttgaat ccattaaggg
     2701 gattgcggtc atggacaagg aaggcaatgt cctgggtcat tccagaattg ctgggacaaa
     2761 ggctgttaga gaaacgctag catccagaat agtgctgttt gggacctcag ctctgattcc
     2821 tgaagtcttc acctactttt ttaaaaggac ccagtatttc aggaaaaacc cagggtcatt
     2881 gtggattttg aaactgtctt gtactgtcct ggcaatggga ctgatggtgc cattttcttt
     2941 tagtatattt ccacagattg gacagataca gtactgtagt cttgaagaga aaattcagtc
     3001 tccaacagaa gaaacagaaa tcttttatca cagaggggtg taggcggccg ctacgtaaat
     3061 tccgcccccc ccccccctct cccycccccc ccctaacgtt actggccgaa gccgcttgga
     3121 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3181 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3241 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3301 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3361 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3421 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3481 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3541 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     3601 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     3661 tggttccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     3721 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     3781 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     3841 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     3901 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     3961 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     4021 aattgctgcc ctctggttat gtgtgggagg gctaagtcga cgataaaata aaagatttta
     4081 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     4141 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     4201 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     4261 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     4321 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4381 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4441 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4501 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     4561 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4621 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4681 agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     4741 ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4801 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5401 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttgcggcc
     5701 ggccgcaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6721 cc
//
