LOCUS       sequence-261586-        6600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged MCART2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  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 6600)
  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-MCART2
FEATURES             Location/Qualifiers
     source          1..6600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      70..79
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             79..144
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(1099..1673,1674..1676,1677..1685)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1266..1283)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1493..1512
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1686..2081
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2252..2721
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2851..2868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3022..3610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3102..3121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3774..4565,4566..4634))
                     /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     4397..4416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4635..4739)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4769..5362
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5425..5782
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5841..6257
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6187..6209
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6225..6241
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(6267..6600,1..41)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     6579..6595
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
ORIGIN
        1 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatcta gctagttaat
       61 taaggatccg ccaccatgga ctacaaggac cacgacggtg actacaagga ccacgacatc
      121 gactacaagg acgacgacga caaggggcat ggcaccggca gcaccggcag cgaattcatg
      181 atagacagtg aggcacatga gaagaggcct ccaattctca caagctctaa acaagacatt
      241 tccccgcaca tcacaaacgt cggggaaatg aaacactatc tatgcggatg ttgcgcggct
      301 ttcaataatg tggcgataac gtacccgata cagaaggtcc tttttcgaca gcaactttac
      361 gggattaaga cgcgcgatgc cgtcctccaa cttcgccggg atggattcag gaatctgtac
      421 cggggcattc tgcctcctct catgcagaag acaacgaccc ttgcgttgat gtttggactg
      481 tatgaggatc tcagttgctt gttgcggaag catgtgcgcg caccagagtt tgccactcac
      541 ggggtcgccg ctgtgctggc agggacggca gaagcaattt tcactcccct cgagagggta
      601 caaacgctgt tgcaaaacca caaacaccac gataaattca caaatactta ccaggctttt
      661 aaagcgttga aatgtcacgg gattggcgaa tattatcgcg gtctcgtacc cattcttttt
      721 cgaaatggat tgagtaacgt cctgtttttt ggtcttcgag ggccgataaa ggagcatctc
      781 ccaactgcaa ctacacacag tgctcatttg gtcaatgatt ttataggtgg aggtctgctc
      841 ggtgctatgc tggggttcct ttgttttcct attaatgttg tcaagacaag gcttcagtca
      901 cagataggag gtgagttcca gtcctttccg aaggtcttcc aaaaaatctg gttggaacgg
      961 gaccgaaaac tcatcaatct gttcagggga gcacatctta actaccatag gagtttgatt
     1021 tcatggggca taatcaacgc aacctatgag ttccttctta aatttattgc ggccgctacg
     1081 taaattccgc cccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg
     1141 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     1201 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     1261 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     1321 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     1381 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     1441 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     1501 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     1561 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     1621 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     1681 caaccatggt tcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     1741 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     1801 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     1861 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     1921 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     1981 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     2041 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agtcgacgat aaaataaaag
     2101 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     2161 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     2221 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     2281 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     2341 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     2401 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     2461 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     2521 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     2581 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     2641 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     2701 cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     2761 aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg
     2821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3181 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3421 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3721 gcggccggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3781 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3841 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3901 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3961 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4021 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4081 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4141 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4201 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4261 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4321 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4381 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4441 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4501 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4561 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4621 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4681 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4741 acatttcccc gaaaagtgcc acctgcataa tgaaagaccc cacctgtagg tttggcaagc
     4801 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagaaaagt
     4861 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     4921 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     4981 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     5041 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     5101 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     5161 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
     5221 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     5281 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     5341 ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg
     5401 accaccgacc caccaccggg aggtaagctg gccagcaact tatctgtgtc tgtccgattg
     5461 tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg
     5521 tatctggcgg acccgtggtg gaactgacga gttcggaaca cccggccgca accctgggag
     5581 acgtcccagg gacttcgggg gccgtttttg tggcccgacc tgagtccaaa aatcccgatc
     5641 gttttggact ctttggtgca cccccctaat aggagggata tgtggttctg gtaggagacg
     5701 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg gaccgaagcc
     5761 gcgccgcgcg tcttgtctgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc
     5821 tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc
     5881 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     5941 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     6001 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     6061 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     6121 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     6181 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     6241 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     6301 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     6361 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     6421 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     6481 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     6541 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
//
