LOCUS       sequence-212922-        6705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Flag-SFXN5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6705)
  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 6705)
  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-SFXN5
FEATURES             Location/Qualifiers
     source          1..6705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             61..654
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    717..1074
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1133..1549
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1479..1501
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1517..1533
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1559..1933
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1871..1887
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1962..1971
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1971..2036
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(2172..2183)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3119..3671
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3263..3280)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3490..3509
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3683..4078
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4249..4718
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4848..4865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5019..5607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5099..5118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5771..6562,6563..6631))
                     /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     6394..6413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6632..6705,1..31))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcat
       61 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
      121 atggaaaaat acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac
      181 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      241 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
      301 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
      361 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
      421 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
      481 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
      541 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
      601 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
      661 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      721 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      781 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      841 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      901 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
      961 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     1021 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
     1081 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1141 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1201 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1261 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1321 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1381 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1441 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1501 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     1561 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     1621 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     1681 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     1741 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     1801 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     1861 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     1921 atcctctaga ctgccggatc tagctagtta attaaggatc cgccaccatg gactacaagg
     1981 accacgacgg tgactacaag gaccacgaca tcgactacaa ggacgacgac gacaaggggc
     2041 atggcaccgg cagcaccggc agcgaattca tggcggatac agcgactaca gcatcggcgg
     2101 cggcggctag tgccgctagc gcctcgagcg atgcacctcc tttccaactg ggcaaacccc
     2161 gcttccagca gacgtccttc tatggccgct tcaggcactt cttggatatc atcgaccctc
     2221 gcacactctt tgtcactgag agacgtctca gagaggctgt gcagctgctg gaggactata
     2281 agcatgggac cctgcgcccg ggggtcacca atgaacagct ctggagtgca cagaaaatca
     2341 agcaggctat tctacatccg gacaccaatg agaagatctt catgccattt agaatgtcag
     2401 gttatattcc ttttgggacg ccaattgtag tcggtcttct cttgcccaac cagacactgg
     2461 catccactgt cttctggcag tggctgaacc agagccacaa tgcctgtgtc aactatgcaa
     2521 accgcaatgc gaccaagcct tcacctgcat ccaagttcat ccagggatac ctgggagctg
     2581 tcatcagcgc tgtctccatt gctgtgggcc ttaatgtcct ggttcagaaa gccaacaagt
     2641 tcaccccagc cacccgcctt ctcatccaga ggtttgtgcc gttccctgct gtagccagtg
     2701 ccaatatctg caatgtggtc ctgatgcggt acggggagct ggaggaaggg attgatgtcc
     2761 tggacagcga tggcaacctc gtgggctcct ccaagatcgc agcccgacac gccctgctgg
     2821 agacggcgct gacgcgagtg gtcctgccca tgcccatcct ggtgctaccc ccgatcgtca
     2881 tgtccatgct ggagaagacg gctctcctgc aggcacgccc ccggctgctc ctccctgtgc
     2941 aaagcctcgt gtgcctggca gccttcggcc tggccctgcc gctggccatc agcctcttcc
     3001 cgcaaatgtc agagattgaa acatcccaat tagagccgga gatagcccag gccacgagca
     3061 gccggacagt ggtgtacaac aaggggttgt gaagcggccg ctacgtaaat tccccctaac
     3121 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     3181 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     3241 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     3301 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     3361 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     3421 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     3481 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     3541 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     3601 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3661 cgatgataat atggccacaa ccatggttcc tttgtctcaa gaagaatcca ccctcattga
     3721 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     3781 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     3841 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     3901 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     3961 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     4021 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt
     4081 cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     4141 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     4201 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     4261 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     4321 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     4381 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     4441 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     4501 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     4561 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     4621 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     4681 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta
     4741 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
     4801 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5701 ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt
     5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgta
//
