LOCUS       sequence-261589-        6583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged MCART1R182A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  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 6583)
  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-MCART1R182A
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             25..441
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     371..393
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     409..425
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    451..825
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     763..779
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      854..863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             863..928
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1873..2439
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1889..2441
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2033..2050)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2260..2279
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2453..2848
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3019..3488
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3618..3635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3789..4377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3869..3888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4541..5332,5333..5401))
                     /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     5164..5183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5402..5506)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5536..6129
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6192..6549
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
       61 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
      121 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
      181 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
      241 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
      301 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
      361 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
      421 ctcactcctt ctctaggcgc ccccatatgg ccatatgaga tcttatatgg ggcacccccg
      481 ccccttgtaa acttccctga ccctgacatg acaagagtta ctaacagccc ctctctccaa
      541 gctcacttac aggctctcta cttagtccag cacgaagtct ggagacctct ggcggcagcc
      601 taccaagaac aactggaccg accggtggta cctcaccctt accgagtcgg cgacacagtg
      661 tgggtccgcc gacaccagac taagaaccta gaacctcgct ggaaaggacc ttacacagtc
      721 ctgctgacca cccccaccgc cctcaaagta gacggcatcg cagcttggat acacgccgcc
      781 cacgtgaagg ctgccgaccc cgggggtgga ccatcctcta gactgccgga tctagctagt
      841 taattaagga tccgccacca tggactacaa ggaccacgac ggtgactaca aggaccacga
      901 catcgactac aaggacgacg acgacaaggg gcatggcacc ggcagcaccg gcagcgaatt
      961 catgatggac tcagaggcgc atgagaaaag gccgccgata ctcaccagct ctaagcagga
     1021 tatctccccg cacataacga atgtcgggga aatgaaacac tatctatgcg gatgctgtgc
     1081 tgcatttaac aatgtagcca ttacgtttcc aatacaaaag gttctcttta ggcaacagct
     1141 ctatggaatt aagacacgag acgcgatatt gcaactccgg cgcgacggat ttagaaacct
     1201 ctacagaggc atccttcctc cactgatgca aaagacaacg acattggccc tcatgtttgg
     1261 tctgtatgag gacctttcct gccttctcca caaacacgtc tcagcgccgg agttcgcaac
     1321 gtccggagtt gcagcagtcc tcgccggcac caccgaagct attttcacgc cgttggagcg
     1381 ggtccaaacg ttgctccaag atcacaaaca tcacgacaaa tttacaaata cataccaagc
     1441 ttttaaagca ctgaagtgtc atggtatcgg tgaatactac cgaggattgg tcccaattct
     1501 tttcgcaaac ggattgagta acgttctgtt cttcgggctt cgcgggccta tcaaagaaca
     1561 ccttcccaca gccacgaccc attctgctca tttggtaaat gatttcattt gtggagggct
     1621 gttgggagcg atgttgggat tcctcttctt tccaataaac gtagtaaaga cccgcattca
     1681 aagccaaatc ggcggtgaat ttcagtcatt cccgaaagtg ttccaaaaga tctggctgga
     1741 acgcgacagg aagctgataa atctgtttcg cggggcgcat cttaactatc accgaagtct
     1801 gataagctgg ggcatcataa atgcaaccta tgagtttttg cttaaagtta tttgagcggc
     1861 cgctacgtaa attccgcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     1921 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     1981 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     2041 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     2101 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     2161 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     2221 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2281 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2341 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     2401 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggttcc
     2461 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     2521 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     2581 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     2641 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     2701 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     2761 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     2821 gccctctggt tatgtgtggg agggctaagt cgacgataaa ataaaagatt ttatttagtc
     2881 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     2941 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     3001 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     3061 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     3121 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca
     3181 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3241 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3301 ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga
     3361 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     3421 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     3481 gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt
     3541 aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3601 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3661 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3721 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3781 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3841 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3901 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3961 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4021 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4081 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4141 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4201 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4261 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4321 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4381 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4441 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccggccgca
     4501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5521 aagtgccacc tgcataatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     5581 gccattttgc aaggcatgga aaaatacata actgagaata gaaaagttca gatcaaggtc
     5641 aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     5701 ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     5761 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag
     5821 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     5881 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     5941 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     6001 tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     6061 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     6121 tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac
     6181 caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
     6241 actgatttta tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc
     6301 cgtggtggaa ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccagggac
     6361 ttcgggggcc gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt
     6421 tggtgcaccc ccctaatagg agggatatgt ggttctggta ggagacgaga acctaaaaca
     6481 gttcccgcct ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct
     6541 tgtctgctgc agcatcgttc tgtgttgtct ctgtctgact gtg
//
