LOCUS       sequence-261723-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged NDT1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  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 6831)
  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-NDT1
FEATURES             Location/Qualifiers
     source          1..6831
                     /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(2178..2752,2753..2755,2756..2764)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2345..2362)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2572..2591
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2765..3160
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3331..3800
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3930..3947)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4101..4689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4181..4200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4853..5644,5645..5713))
                     /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     5476..5495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5714..5818)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5848..6441
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    join(6504..6831,1..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 aattcatgac ccaaaccgat aaccccgttc cgaactgtgg tctgttgccc gagcagcagt
     1081 attgctcagc agatcacgaa gagcctcttc tgctccatga agaacaattg atttttccag
     1141 accattcctc acaacttagt tcagcggaca ttattgaacc gatcaagatg aactcttcaa
     1201 cagaatcaat cattgggacg acgcttcgga agaaatgggt tcccctcagc agcacgcaaa
     1261 tcaccgccct ctccggggct tttgcagggt ttctcagcgg cgtggctgta tgccccctgg
     1321 acgtggcgaa aaccagattg caggcgcaag gcttgcagac tcggttcgaa aacccctatt
     1381 acagaggcat catggggaca ctttctacta ttgtacgaga tgaaggcccc cggggactgt
     1441 acaagggtct ggtccctata gttcttggtt atttccccac atggatgatc tatttctctg
     1501 tttatgagtt ctccaagaaa ttttttcacg gcatcttccc gcaattcgac ttcgttgccc
     1561 aatcatgcgc tgcgataacg gcaggagcag cgtctactac gctcacaaac ccaatatggg
     1621 tagtaaaaac tcgattgatg ctgcaaagta atctgggcga gcatccgact cattacaagg
     1681 gaaccttcga cgcttttcgg aagctcttct accaagaggg attcaaagcc ttgtacgccg
     1741 ggcttgtgcc aagccttctc ggactgttcc atgttgctat tcattttcct atttacgagg
     1801 accttaaagt gcgcttccat tgttattcac gcgagaacaa cactaacagc ataaacctcc
     1861 agagactgat aatggcatcc agtgtctcaa agatgattgc cagtgcggtg acgtatcccc
     1921 acgaaatact ccgcactcgg atgcaactca agtctgacat acccgactca atacagaggc
     1981 gcttgttccc cctcataaaa gcgacctatg ctcaggaagg cctgaaggga ttttacagtg
     2041 gtttcaccac caaccttgtt cgcaccatac ccgcttctgc aattactttg gttagctttg
     2101 agtattttag aaacaggttg gagaatattt caacgatggt tatctaagcg gccgctacgt
     2161 aaattccgcc cccccccccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     2221 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     2281 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     2341 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     2401 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     2461 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     2521 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     2581 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     2641 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     2701 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
     2761 aaccatggtt cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     2821 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     2881 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     2941 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     3001 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     3061 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     3121 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa gtcgacgata aaataaaaga
     3181 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     3241 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     3301 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     3361 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     3421 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     3481 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     3541 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     3601 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
     3661 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     3721 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     3781 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     3841 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     3901 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3961 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4021 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4501 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4741 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     4801 cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4861 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4921 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4981 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5041 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5101 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5161 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5221 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5281 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5341 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5401 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5461 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5521 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5581 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5641 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5701 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5761 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5821 catttccccg aaaagtgcca cctgcataat gaaagacccc acctgtaggt ttggcaagct
     5881 agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa tagaaaagtt
     5941 cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     6001 aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa
     6061 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
     6121 gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
     6181 ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg
     6241 ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc
     6301 cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt
     6361 tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
     6421 cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc tgcccaggga
     6481 ccaccgaccc accaccggga ggtaagctgg ccagcaactt atctgtgtct gtccgattgt
     6541 ctagtgtcta tgactgattt tatgcgcctg cgtcggtact agttagctaa ctagctctgt
     6601 atctggcgga cccgtggtgg aactgacgag ttcggaacac ccggccgcaa ccctgggaga
     6661 cgtcccaggg acttcggggg ccgtttttgt ggcccgacct gagtccaaaa atcccgatcg
     6721 ttttggactc tttggtgcac ccccctaata ggagggatat gtggttctgg taggagacga
     6781 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttggg a
//
