LOCUS       sequence-267545-        6630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged MCART6 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6630)
  AUTHORS   Kory N, Uit de Bos J, van der Rijt S, Jankovic N, Gura M, Arp N, 
            Pena IA, Prakash G, Chan SH, Kunchok T, Lewis CA, Sabatini DM
  TITLE     MCART1/SLC25A51 is required for mitochondrial NAD transport.
  JOURNAL   Sci Adv. 2020 Oct 21;6(43):eabe5310. doi: 10.1126/sciadv.abe5310. 
            Print 2020 Oct.
  PUBMED    33087354
REFERENCE   2  (bases 1 to 6630)
  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: "Sci Adv."; 
            date: "2020-10-21"; volume: "6(43):eabe5310. doi"; pages: " 
            10.1126/sciadv.abe5310. Print 2020 Oct"
COMMENT     Sequence Label: pMXs_FLAG-MCART6
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             58..474
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     404..426
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     442..458
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    484..858
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     796..812
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      887..896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             896..961
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(1946..2520,2521..2523,2524..2532)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2113..2130)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2340..2359
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2533..2928
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3099..3568
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3698..3715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3869..4457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3949..3968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4621..5412,5413..5481))
                     /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     5244..5263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5482..5586)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5616..6209
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6272..6629
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc
       61 cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc
      121 gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa
      181 tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc
      241 caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac
      301 atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac
      361 cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt
      421 tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg cgcccccata
      481 tggccatatg agatcttata tggggcaccc ccgccccttg taaacttccc tgaccctgac
      541 atgacaagag ttactaacag cccctctctc caagctcact tacaggctct ctacttagtc
      601 cagcacgaag tctggagacc tctggcggca gcctaccaag aacaactgga ccgaccggtg
      661 gtacctcacc cttaccgagt cggcgacaca gtgtgggtcc gccgacacca gactaagaac
      721 ctagaacctc gctggaaagg accttacaca gtcctgctga ccacccccac cgccctcaaa
      781 gtagacggca tcgcagcttg gatacacgcc gcccacgtga aggctgccga ccccgggggt
      841 ggaccatcct ctagactgcc ggatctagct agttaattaa ggatccgcca ccatggacta
      901 caaggaccac gacggtgact acaaggacca cgacatcgac tacaaggacg acgacgacaa
      961 ggggcatggc accggcagca ccggcagcga attcatgggc gagcaaaatc atagcccggg
     1021 caaggagctt caacatcgga cacgcgctga ggctcctggg aagaaatcct ggcattcaca
     1081 agcctacgca cttggcgcag tctctaactt tatgagtacg tttttgacct tcccaatata
     1141 taaggtggtc ttcaggcaac aaatacatgc tatggcggtc tccgaagcgg taagacagct
     1201 gtggcatgag ggtccacagt atttttatag gggcatctat ccccctctgc tttctaaaac
     1261 acttcaaggc accttgcttt ttggaacgta tgacagcctc ttgtgcttcc tttcaccagt
     1321 tggaccgcat accctgggtc atagatgggc agccggattg atgtccgggg ttgtggaggc
     1381 ggtagccctt agtccgtttg aacgggtaca aaacgtgctt caagatggcc gcaaacaagc
     1441 acgattccct agcactttct ctattctgaa agaatttaac tcatatggcc tttggggaag
     1501 gctgtctctc ggttattatc gggggttctg gcccgtactc gcaagaaatt cacttggctc
     1561 agctctctac ttctccttta aagaccccat acaggacggg ttggcagagc aagggttgcc
     1621 gcattgggtc ccggcgctgg tgagtgggag cgttaacggc actataactt gcctcgttct
     1681 ttaccctttg atcgtgttgg tggcgaatat gcaaagccac atcggctggc agaacatgcc
     1741 ctctctttgg gcgagcgccc aggacgtttg gaacactcga ggtcgcaagc tgctgcttat
     1801 ctataggggt ggctctctcg taatactcag gtctagtgtg acttggggcc ttacgaccgc
     1861 gatccacgac tttctccaga ggaagtctca ttctagaaag gagctcaaga cggatgcggc
     1921 cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac gttactggcc
     1981 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2041 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     2101 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2161 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2221 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2281 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2341 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2401 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     2461 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2521 atggccacaa ccatggttcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     2581 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     2641 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     2701 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     2761 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     2821 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     2881 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt cgacgataaa
     2941 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     3001 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     3061 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     3121 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     3181 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     3241 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     3301 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     3361 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     3421 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     3481 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     3541 ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt
     3601 ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg
     3661 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3721 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3781 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3841 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3901 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3961 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4021 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4081 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4141 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4201 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4261 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4321 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4381 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4441 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4501 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4561 tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4621 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4681 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4741 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4801 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4861 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4921 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4981 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5041 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5101 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5161 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5221 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5281 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5341 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5401 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5461 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5521 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5581 tccgcgcaca tttccccgaa aagtgccacc tgcataatga aagaccccac ctgtaggttt
     5641 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     5701 gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     5761 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     5821 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     5881 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     5941 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     6001 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     6061 cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     6121 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     6181 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
     6241 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
     6301 ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact
     6361 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc
     6421 ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga gtccaaaaat
     6481 cccgatcgtt ttggactctt tggtgcaccc ccctaatagg agggatatgt ggttctggta
     6541 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttgggac
     6601 cgaagccgcg ccgcgcgtct tgtctgctgc
//
