LOCUS       sequence-261715-        6717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged NDT2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6717)
  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 6717)
  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-NDT2
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             89..505
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     435..457
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     473..489
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    515..889
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     827..843
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             927..992
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(2064..2638,2639..2641,2642..2650)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2231..2248)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2458..2477
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2651..3046
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3217..3686
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3816..3833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4739..5530,5531..5599))
                     /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     5362..5381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5600..5704)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5734..6327
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    join(6390..6717,1..30)
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc tctgtctgac
       61 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
      121 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
      181 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
      241 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
      301 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
      361 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
      421 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
      481 agccctcact ccttctctag gcgcccccat atggccatat gagatcttat atggggcacc
      541 cccgcccctt gtaaacttcc ctgaccctga catgacaaga gttactaaca gcccctctct
      601 ccaagctcac ttacaggctc tctacttagt ccagcacgaa gtctggagac ctctggcggc
      661 agcctaccaa gaacaactgg accgaccggt ggtacctcac ccttaccgag tcggcgacac
      721 agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag gaccttacac
      781 agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt ggatacacgc
      841 cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc cggatctagc
      901 tagttaatta aggatccgcc accatggact acaaggacca cgacggtgac tacaaggacc
      961 acgacatcga ctacaaggac gacgacgaca aggggcatgg caccggcagc accggcagcg
     1021 aattttcaag ggtagtttta ttatccccat tgttcatgaa ttctaaaaat gcaagcctgg
     1081 cgaacggtaa ttatgctatc cccacgaagt tgaataggct taagaagaac gcggacccac
     1141 gagtcgcagc aataagtgga gccctcagcg gtgcccttag cgcgatgctc gtttgccctt
     1201 ttgatgttgc taaaacccgc cttcaagcgc aagggctcca aaacatgact caccaaagcc
     1261 agcattataa agggttcttc ggcaccttcg ccactatatt taaagatgaa ggcgcggcgg
     1321 gcttgtacaa agggctgcaa cccaccgtct tgggctatat acctactctc atgatttatt
     1381 tctctgttta cgatttttgt cggaaatact cagtcgacat ctttcctcat agcccttttt
     1441 tgagtaacgc atcttcagcg ataacagccg gggccatttc cacggttgcg accaatccaa
     1501 tttgggtcgt aaaaacacgc cttatgctgc agaccggaat cggcaagtat agcacgcact
     1561 acaaaggtac tattgatacg ttcaggaaga ttattcaaca agagggcgct aaggccctgt
     1621 acgcgggtct cgtgcctgca ctcctgggga tgcttaacgt cgctatacaa tttcctcttt
     1681 atgaaaatct taagatacgc tttggttatt cagagagtac ggacgtcagt acagatgtga
     1741 cctccagtaa tttccaaaag ctgattctcg catcaatgct ctctaagatg gtagcctcta
     1801 ctgtgactta cccgcatgaa attctgagaa cgagaatgca gcttaagtct gatcttccga
     1861 atacagtaca gcggcatttg ttgcccttga taaaaattac ctaccgacaa gaaggattcg
     1921 caggtttcta ttctggattt gcaacaaacc ttgtcagaac cgtacctgcc gccgtagtaa
     1981 cactcgtgtc tttcgaatat agcaaaaaat accttaccac attctttcag taagcggccg
     2041 ctacgtaaat tccgcccccc ccccccctct ccctcccccc cccctaacgt tactggccga
     2101 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     2161 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     2221 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     2281 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     2341 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     2401 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     2461 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     2521 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     2581 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     2641 ggccacaacc atggttcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     2701 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     2761 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     2821 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     2881 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     2941 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     3001 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagtcg acgataaaat
     3061 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     3121 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag
     3181 agaagttcag atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat
     3241 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg
     3301 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     3361 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
     3421 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3481 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3541 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
     3601 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     3661 gactacccgt cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt
     3721 ttcttaagta tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg
     3781 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4681 cttttgcggc cggccgcaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4741 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4801 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4861 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4921 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4981 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5041 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5101 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5161 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5221 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5281 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5341 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5401 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5461 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5521 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5581 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5641 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5701 cgcgcacatt tccccgaaaa gtgccacctg cataatgaaa gaccccacct gtaggtttgg
     5761 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     5821 aaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     5881 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     5941 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     6001 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     6061 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     6121 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     6181 gcgcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     6241 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     6301 actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga gacccctgcc
     6361 cagggaccac cgacccacca ccgggaggta agctggccag caacttatct gtgtctgtcc
     6421 gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt agctaactag
     6481 ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg ccgcaaccct
     6541 gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt ccaaaaatcc
     6601 cgatcgtttt ggactctttg gtgcaccccc ctaataggag ggatatgtgg ttctggtagg
     6661 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggga
//
