LOCUS       sequence-314032-        6693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of flag-tagged yeast FSF1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  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 6693)
  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-FSF1
FEATURES             Location/Qualifiers
     source          1..6693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             46..639
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    702..1059
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1118..1534
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1464..1486
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1502..1518
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1544..1918
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1856..1872
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1947..1956
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1956..2021
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(3069..3643,3644..3646,3647..3655)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3236..3253)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3463..3482
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3656..4051
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4222..4691
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4821..4838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4992..5580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5072..5091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5744..6535,6536..6604))
                     /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     6367..6386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6605..6693,1..16))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga aagaccccac
       61 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
      121 actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
      181 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
      241 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
      301 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
      361 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
      421 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
      481 acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
      541 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
      601 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
      661 gagacccctg cccagggacc accgacccac caccgggagg taagctggcc agcaacttat
      721 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
      781 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc
      841 ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga
      901 gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc ccctaatagg agggatatgt
      961 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     1021 ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct
     1081 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
     1141 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
     1201 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
     1261 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
     1321 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
     1381 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
     1441 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
     1501 cctttatcca gccctcactc cttctctagg cgcccccata tggccatatg agatcttata
     1561 tggggcaccc ccgccccttg taaacttccc tgaccctgac atgacaagag ttactaacag
     1621 cccctctctc caagctcact tacaggctct ctacttagtc cagcacgaag tctggagacc
     1681 tctggcggca gcctaccaag aacaactgga ccgaccggtg gtacctcacc cttaccgagt
     1741 cggcgacaca gtgtgggtcc gccgacacca gactaagaac ctagaacctc gctggaaagg
     1801 accttacaca gtcctgctga ccacccccac cgccctcaaa gtagacggca tcgcagcttg
     1861 gatacacgcc gcccacgtga aggctgccga ccccgggggt ggaccatcct ctagactgcc
     1921 ggatctagct agttaattaa ggatccgcca ccatggacta caaggaccac gacggtgact
     1981 acaaggacca cgacatcgac tacaaggacg acgacgacaa ggggcatggc accggcagca
     2041 ccggcagcga attcatggcg tctagcgttc caggcccgat tgaccttcca gagagcaggt
     2101 atgacctttc tacttactgg ggccggatac gacattgtgc tgagattagc gatcccacaa
     2161 tgttgctcac cacagaaaaa gatctcgctc acgcaaggga gataatttct gcatatcggc
     2221 acggggaact gaaagaaaca acacccgagt tttggagagc gaagaagcag cttgactcca
     2281 cagtgcatcc cgatacaggg aagacggtac tgctcccctt tagaatgtct agcaatgttc
     2341 tgtctaatct tgttgtgaca gttggtatgc tcactccagg gctgggcacg gcgggaaccg
     2401 tgttttggca gtgggcaaat cagtcactca atgtagccgt taacagtgca aacgctaata
     2461 aaagccatcc tatgtctact tctcaacttc tcactaatta cgctgctgca gtcactgcgt
     2521 cctgcggtgt cgcattgggg ctgaataact tggttcctcg cctgaagaac atctctccac
     2581 attccaaact tatactgggc cgcctcgttc ccttcgctgc agtcgtaagc gccggtatag
     2641 taaatgtatt cctgatgagg ggcaacgaga tccggaaagg tataagcgtc tttgattcta
     2701 acggtgatga agtgggaaag agtaagaagg cggccttcat ggcggtcgga gagactgctc
     2761 ttagtcgggt gattaatgct acaccaacaa tggtgattcc tccactgata ctggtccgcc
     2821 tgcagcgagg tgtgcttaag ggaaagtctc ttggtgtcca gactctcgcc aatctgggac
     2881 tgatctccgt gacgatgttt agtgccttgc cattcgccct tggcattttc ccgcaacgcc
     2941 aagcgatcca cttgaataaa ctcgaacccg aactgcatgg caagaaggac aaagatggaa
     3001 agcctatcga gaaggtttat ttcaaccgag gaatttaagc ggccgctacg taaattccgc
     3061 cccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     3121 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3181 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3241 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3301 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     3361 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     3421 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3481 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     3541 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     3601 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
     3661 tcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     3721 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     3781 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     3841 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     3901 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     3961 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     4021 gctgccctct ggttatgtgt gggagggcta agtcgacgat aaaataaaag attttattta
     4081 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4141 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     4201 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     4261 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     4321 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     4381 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     4441 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     4501 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     4561 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     4621 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     4681 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     4741 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4801 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4861 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4921 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4981 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5041 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5101 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5161 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5221 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5281 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5341 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5401 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5461 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5521 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5581 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5641 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
     5701 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6661 gatacatatt tgaatgtatt tagaaaaata aac
//
