LOCUS       sequence-273755-        6603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged MCART1Q90A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6603)
  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 6603)
  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-MCART1Q90A
FEATURES             Location/Qualifiers
     source          1..6603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             39..455
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     385..407
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     423..439
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    465..839
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     777..793
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      868..877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             877..942
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(1900..2474,2475..2477,2478..2486)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2067..2084)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2294..2313
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2487..2882
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3053..3522
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3652..3669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3823..4411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3903..3922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4575..5366,5367..5435))
                     /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     5198..5217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5436..5540)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5570..6163
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6226..6583
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
       61 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
      121 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
      181 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
      241 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
      301 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
      361 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
      421 ccctttatcc agccctcact ccttctctag gcgcccccat atggccatat gagatcttat
      481 atggggcacc cccgcccctt gtaaacttcc ctgaccctga catgacaaga gttactaaca
      541 gcccctctct ccaagctcac ttacaggctc tctacttagt ccagcacgaa gtctggagac
      601 ctctggcggc agcctaccaa gaacaactgg accgaccggt ggtacctcac ccttaccgag
      661 tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag
      721 gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt
      781 ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc
      841 cggatctagc tagttaatta aggatccgcc accatggact acaaggacca cgacggtgac
      901 tacaaggacc acgacatcga ctacaaggac gacgacgaca aggggcatgg caccggcagc
      961 accggcagcg aattcatgat ggactcagag gcgcatgaga aaaggccgcc gatactcacc
     1021 agctctaagc aggatatctc cccgcacata acgaatgtcg gggaaatgaa acactatcta
     1081 tgcggatgct gtgctgcatt taacaatgta gccattacgt ttccaataca aaaggttctc
     1141 tttaggcaac agctctatgg aattaagaca cgagacgcga tattgcaact ccggcgcgac
     1201 ggatttagaa acctctacag aggcatcctt cctccactga tggcaaagac aacgacattg
     1261 gccctcatgt ttggtctgta tgaggacctt tcctgccttc tccacaaaca cgtctcagcg
     1321 ccggagttcg caacgtccgg agttgcagca gtcctcgccg gcaccaccga agctattttc
     1381 acgccgttgg agcgggtcca aacgttgctc caagatcaca aacatcacga caaatttaca
     1441 aatacatacc aagcttttaa agcactgaag tgtcatggta tcggtgaata ctaccgagga
     1501 ttggtcccaa ttcttttcag aaacggattg agtaacgttc tgttcttcgg gcttcgcggg
     1561 cctatcaaag aacaccttcc cactgccacc acacacagcg ctcatttggt aaatgatttc
     1621 atttgtggag ggctgttggg agcgatgttg ggattcctct tctttccaat aaacgtagta
     1681 aagacccgca ttcaaagcca aatcggcggt gaatttcagt cattcccgaa agtgttccaa
     1741 aagatctggc tggaacgcga caggaagctg ataaatctgt ttcgcggggc gcatcttaac
     1801 tatcaccgaa gtctgataag ctggggcatc ataaatgcaa cctatgagtt tttgcttaaa
     1861 gttatttgag cggccgctac gtaaattccg cccccccccc ccctctccct cccccccccc
     1921 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     1981 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2041 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2101 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2161 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2221 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2281 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2341 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     2401 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     2461 aacacgatga taatatggcc acaaccatgg ttcctttgtc tcaagaagaa tccaccctca
     2521 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
     2581 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
     2641 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
     2701 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
     2761 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
     2821 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
     2881 aagtcgacga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     2941 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaaat
     3001 acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac agctgaatat
     3061 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     3121 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     3181 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
     3241 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     3301 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     3361 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
     3421 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
     3481 tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa
     3541 attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat
     3601 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3661 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3721 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3781 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3841 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3901 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3961 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4021 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4081 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4141 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4201 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4261 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4321 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4381 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4441 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4501 gatcttcacc tagatccttt tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa
     4561 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4621 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4681 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4741 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4801 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4861 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4921 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4981 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5041 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5101 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5161 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5221 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5281 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5341 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5401 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5461 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5521 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgcata atgaaagacc
     5581 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     5641 cataactgag aatagaaaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     5701 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     5761 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     5821 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     5881 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     5941 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     6001 cacaacccct cactcggcgc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     6061 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     6121 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga
     6181 tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac
     6241 ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta
     6301 ctagttagct aactagctct gtatctggcg gacccgtggt ggaactgacg agttcggaac
     6361 acccggccgc aaccctggga gacgtcccag ggacttcggg ggccgttttt gtggcccgac
     6421 ctgagtccaa aaatcccgat cgttttggac tctttggtgc acccccctaa taggagggat
     6481 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     6541 tttcggtttg ggaccgaagc cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt
     6601 gtc
//
