LOCUS       sequence-261721-        6594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged MCART1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  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 6594)
  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-MCART1
FEATURES             Location/Qualifiers
     source          1..6594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1923..1932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1932..1997
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2969..3521
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3113..3130)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3340..3359
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3533..3928
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4099..4568
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4698..4715)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4869..5457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4949..4968)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5621..6412,6413..6481))
                     /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     6244..6263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6482..6586)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 ccgccaccat ggactacaag gaccacgacg gtgactacaa ggaccacgac atcgactaca
     1981 aggacgacga cgacaagggg catggcaccg gcagcaccgg cagcgaattc atgatggact
     2041 cagaggcgca tgagaaaagg ccgccgatac tcaccagctc taagcaggat atctccccgc
     2101 acataacgaa tgtcggggaa atgaaacact atctatgcgg atgctgtgct gcatttaaca
     2161 atgtagccat tacgtttcca atacaaaagg ttctctttag gcaacagctc tatggaatta
     2221 agacacgaga cgcgatattg caactccggc gcgacggatt tagaaacctc tacagaggca
     2281 tccttcctcc actgatgcaa aagacaacga cattggccct catgtttggt ctgtatgagg
     2341 acctttcctg ccttctccac aaacacgtct cagcgccgga gttcgcaacg tccggagttg
     2401 cagcagtcct cgccggcacc accgaagcta ttttcacgcc gttggagcgg gtccaaacgt
     2461 tgctccaaga tcacaaacat cacgacaaat ttacaaatac ataccaagct tttaaagcac
     2521 tgaagtgtca tggtatcggt gaatactacc gaggattggt cccaattctt ttcagaaacg
     2581 gattgagtaa cgttctgttc ttcgggcttc gcgggcctat caaagaacac cttcccacag
     2641 ccacgaccca ttctgctcat ttggtaaatg atttcatttg tggagggctg ttgggagcga
     2701 tgttgggatt cctcttcttt ccaataaacg tagtaaagac ccgcattcaa agccaaatcg
     2761 gcggtgaatt tcagtcattc ccgaaagtgt tccaaaagat ctggctggaa cgcgacagga
     2821 agctgataaa tctgtttcgc ggggcgcatc ttaactatca ccgaagtctg ataagctggg
     2881 gcatcataaa tgcaacctat gagtttttgc ttaaagttat ttgagcggcc gctacgtaaa
     2941 ttccgccccc cccccycccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     3001 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     3061 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     3121 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     3181 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     3241 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     3301 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     3361 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     3421 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     3481 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggttcc
     3541 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     3601 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     3661 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     3721 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     3781 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     3841 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     3901 gccctctggt tatgtgtggg agggctaagt cgacgataaa ataaaagatt ttatttagtc
     3961 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     4021 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     4081 caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct
     4141 gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg
     4201 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca
     4261 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     4321 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     4381 ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga
     4441 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     4501 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     4561 gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt
     4621 aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4681 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4741 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4801 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4861 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4921 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4981 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5041 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5101 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5161 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5221 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5281 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5341 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5401 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5461 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5521 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccggccgca
     5581 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5641 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5701 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5761 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5821 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5881 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5941 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6001 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6061 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6121 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6181 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6241 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6301 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6361 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6421 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6481 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6541 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttc
//
