LOCUS       sequence-261676-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged MCART1K91A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6584)
  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 6584)
  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-MCART1K91A
FEATURES             Location/Qualifiers
     source          1..6584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    587..944
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1003..1419
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1349..1371
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1387..1403
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1429..1803
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1741..1757
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1832..1841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1841..1906
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2851..3418
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2868..3420
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3012..3029)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3239..3258
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3432..3827
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3998..4467
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4597..4614)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4768..5356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4848..4867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5520..6311,6312..6380))
                     /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     6143..6162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6381..6485)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(6515..6584,1..524)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 acataactga gaatagaaaa gttcagatca aggtcaggaa cagatggaac agctgaatat
       61 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
      121 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
      181 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
      241 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
      301 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
      361 ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
      421 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
      481 tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg
      541 atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      601 cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttcggaa
      721 cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt tgtggcccga
      781 cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta ataggaggga
      841 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
      901 ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt
      961 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
     1021 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     1081 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     1141 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     1201 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
     1261 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
     1321 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
     1381 tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc
     1441 ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact
     1501 aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg
     1561 agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac
     1621 cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg
     1681 aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca
     1741 gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctaga
     1801 ctgccggatc tagctagtta attaaggatc cgccaccatg gactacaagg accacgacgg
     1861 tgactacaag gaccacgaca tcgactacaa ggacgacgac gacaaggggc atggcaccgg
     1921 cagcaccggc agcgaattca tgatggactc agaggcgcat gagaaaaggc cgccgatact
     1981 caccagctct aagcaggata tctccccgca cataacgaat gtcggggaaa tgaaacacta
     2041 tctatgcgga tgctgtgctg catttaacaa tgtagccatt acgtttccaa tacaaaaggt
     2101 tctctttagg caacagctct atggaattaa gacacgagac gcgatattgc aactccggcg
     2161 cgacggattt agaaacctct acagaggcat ccttcctcca ctgatgcaag cgacaacgac
     2221 attggccctc atgtttggtc tgtatgagga cctttcctgc cttctccaca aacacgtctc
     2281 agcgccggag ttcgcaacgt ccggagttgc agcagtcctc gccggcacca ccgaagctat
     2341 tttcacgccg ttggagcggg tccaaacgtt gctccaagat cacaaacatc acgacaaatt
     2401 tacaaataca taccaagctt ttaaagcact gaagtgtcat ggtatcggtg aatactaccg
     2461 aggattggtc ccaattcttt tcagaaacgg attgagtaac gttctgttct tcgggcttcg
     2521 cgggcctatc aaagaacacc ttcccacagc cacgacccat tctgctcatt tggtaaatga
     2581 tttcatttgt ggagggctgt tgggagcgat gttgggattc ctcttctttc caataaacgt
     2641 agtaaagacc cgcattcaaa gccaaatcgg cggtgaattt cagtcattcc cgaaagtgtt
     2701 ccaaaagatc tggctggaac gcgacaggaa gctgataaat ctgtttcgcg gggcgcatct
     2761 taactatcac cgaagtctga taagctgggg catcataaat gcaacctatg agtttttgct
     2821 taaagttatt tgagcggccg ctacgtaaat tccgcccccc ccccctaacg ttactggccg
     2881 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     2941 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     3001 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     3061 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     3121 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     3181 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     3241 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     3301 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     3361 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     3421 tggccacaac catggttcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
     3481 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
     3541 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
     3601 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
     3661 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
     3721 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
     3781 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagtc gacgataaaa
     3841 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     3901 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     3961 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     4021 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     4081 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4141 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     4201 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4261 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4321 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     4381 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4441 tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt
     4501 tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg
     4561 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4621 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4681 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4741 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4801 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4861 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4921 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4981 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5041 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5101 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5161 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5221 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5281 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5341 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5401 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5461 ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5821 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6121 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6481 ccgcgcacat ttccccgaaa agtgccacct gcataatgaa agaccccacc tgtaggtttg
     6541 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaat
//
