LOCUS       sequence-217244-        6670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Flag-SFXN3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6670)
  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 6670)
  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-SFXN3
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             35..628
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    691..1048
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1107..1523
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1453..1475
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1491..1507
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1533..1907
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1845..1861
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1936..1945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1945..2010
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3039..3608
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3058..3610
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3202..3219)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3429..3448
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3622..4017
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4188..4657
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4787..4804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4958..5546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5038..5057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5710..6501,6502..6570))
                     /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     6333..6352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6571..6670,1..5))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgcgcacat ttccccgaaa agtgccacct gcataatgaa agaccccacc tgtaggtttg
       61 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
      121 aaaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
      181 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
      241 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
      301 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
      361 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
      421 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
      481 ggcgcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
      541 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
      601 gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc
      661 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      721 cgattgtcta gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta
      781 gctctgtatc tggcggaccc gtggtggaac tgacgagttc ggaacacccg gccgcaaccc
      841 tgggagacgt cccagggact tcgggggccg tttttgtggc ccgacctgag tccaaaaatc
      901 ccgatcgttt tggactcttt ggtgcacccc cctaatagga gggatatgtg gttctggtag
      961 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc
     1021 gaagccgcgc cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc tgtctgactg
     1081 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1141 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1201 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1261 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1321 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1381 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1441 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1501 ccctcactcc ttctctaggc gcccccatat ggccatatga gatcttatat ggggcacccc
     1561 cgccccttgt aaacttccct gaccctgaca tgacaagagt tactaacagc ccctctctcc
     1621 aagctcactt acaggctctc tacttagtcc agcacgaagt ctggagacct ctggcggcag
     1681 cctaccaaga acaactggac cgaccggtgg tacctcaccc ttaccgagtc ggcgacacag
     1741 tgtgggtccg ccgacaccag actaagaacc tagaacctcg ctggaaagga ccttacacag
     1801 tcctgctgac cacccccacc gccctcaaag tagacggcat cgcagcttgg atacacgccg
     1861 cccacgtgaa ggctgccgac cccgggggtg gaccatcctc tagactgccg gatctagcta
     1921 gttaattaag gatccgccac catggactac aaggaccacg acggtgacta caaggaccac
     1981 gacatcgact acaaggacga cgacgacaag gggcatggca ccggcagcac cggcagcgaa
     2041 ttcatggaaa gcaaaatggg tgaattgcct ttagacatca acatccagga acctcgctgg
     2101 gaccaaagta ctttcctggg cagagcccgg cactttttca ctgttactga tcctcgaaat
     2161 ctgctgctgt ccggggcaca gctggaagct tctcggaaca tcgtgcagaa ctacagggcc
     2221 ggcgtggtga ccccagggat caccgaggac cagctgtgga gggccaagta tgtgtatgac
     2281 tccgccttcc atccggacac aggggagaag gtggtcctga ttggccgcat gtcagcccag
     2341 gtgcccatga acatgaccat cactggctgc atgctcacat tctacaggaa gaccccaacc
     2401 gtggtgttct ggcagtgggt gaatcagtcc ttcaatgcca ttgttaacta ctccaaccgc
     2461 agtggtgaca ctcccatcac tgtgaggcag ctggggacag cctatgtgag tgccaccact
     2521 ggagctgtgg ccacggccct gggactcaaa tccctcacca agcacctgcc ccccttggtc
     2581 ggcagatttg tgccctttgc agcagtggca gctgccaact gcatcaacat ccccctgatg
     2641 aggcagagag agctgcaggt gggcatcccg gtggctgatg aggcaggtca gaggcttggc
     2701 tactcggtga ctgcagccaa gcagggaatc ttccaggtgg tgatttcaag aatctgcatg
     2761 gcgattcctg ccatggccat cccaccactg atcatggaca ctctggagaa gaaagacttc
     2821 ctgaagcgcc gcccctggct gggggcaccc ctgcaggtgg gactggtggg cttctgcctg
     2881 gtatttgcaa cccccctgtg ctgtgcccta ttcccccaga agagctccat acacataagc
     2941 aacctggaac cagagctgag agctcagatc catgagcaaa accccagcgt tgaagtggtc
     3001 tactacaaca aggggctttg agcggccgct acgtaaattc cgcccccccc ccccctaacg
     3061 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3121 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3181 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3241 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     3301 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3361 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3421 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3481 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3541 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3601 gatgataata tggccacaac catggttcct ttgtctcaag aagaatccac cctcattgaa
     3661 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     3721 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     3781 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     3841 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     3901 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     3961 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagtc
     4021 gacgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     4081 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     4141 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     4201 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     4261 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     4321 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     4381 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     4441 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4501 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     4561 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     4621 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     4681 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     4741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5641 tcacctagat ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg
     5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt
//
