LOCUS       sequence-261579-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged NMNAT3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6738)
  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 6738)
  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_NMNAT3-FLAG
FEATURES             Location/Qualifiers
     source          1..6738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..420
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     350..372
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     388..404
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    430..804
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     742..758
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1910..1975
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     gap             1994..2004
                     /estimated_length=11
     misc_feature    2006..2573
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2023..2575
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2167..2184)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2394..2413
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2587..2982
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3153..3622
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3752..3769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3923..4511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4003..4022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4675..5466,5467..5535))
                     /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     5298..5317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5536..5640)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5670..6263
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6326..6683
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
       61 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
      121 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
      181 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
      241 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
      301 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
      361 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
      421 cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa cttccctgac
      481 cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca ggctctctac
      541 ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca actggaccga
      601 ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg acaccagact
      661 aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac ccccaccgcc
      721 ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc tgccgacccc
      781 gggggtggac catcctctag actgccggat ctagctagtt aattaaggat ccgccaccat
      841 gaagagccga atacctgtgg tgctcctggc ctgtggctcc tttaacccca tcaccaacat
      901 gcacctgcgc atgtttgagg tggccagaga tcacctacac caaacagaat atgaagaaaa
      961 ggaccatgaa gttgcacttg tttttcctgg acctcagtct atctcaataa aagatatttc
     1021 ttttcatctg caaaaaagga ttcaaaataa tgttagcagc aaaaatgatg accctgacaa
     1081 gtcatctttt aaacgcaaaa gaactgaaga acaagcgttc tgtgatttga atgacagcaa
     1141 gtgcaaaggc acaacactga aaaaaattat atttatagat agcacctgga accaaacaaa
     1201 caaaatattc actgatgagc gacttcaagg aatgtaccag gtcatccagg gtatcatctc
     1261 tcctgtcaac gacacctatg ggaagaaaga cctcgcagct tctcatcacc gagtggccat
     1321 ggcccggctg gccctgcaga catccgactg gatacgggtg gacccttggg agagtgagca
     1381 ggcacagtgg atggagacag tgaaggtgct gaggcatcat cacagcaaac tgctcagatc
     1441 tccaccccag atggaaggcc cagaccatgg caaggcactc ttctcgaccc ctgcagctgt
     1501 gcctgagctg aagcttctct gtggggcaga cgtcttgaag accttccaga cccccaacct
     1561 ctggaaggat gcgcacatcc aggaaatagt ggagaagttt ggcttggtgt gcgtgggccg
     1621 agtaggtcac gacccaaaag gttacatcgc agaatctccc atcctacgga tgcaccagca
     1681 caacattcac ctggccaagg agcctgtgca gaatgagatc agtgccacat acatcaggcg
     1741 agccttgggc caagggcaga gcgtaaagta cctgattccc gatgctgtca tcacgtacat
     1801 caaggaccat ggcctctaca ccaagggcag tacctggaaa ggcaaaagca cccagagcac
     1861 tgagggcaag acaagcgcgg ccgcaggagg cggtggaggc ggtgcaggcg actacaaaga
     1921 ccatgacggt gattataaag atcatgacat tgattacaag gatgacgatg acaagtaata
     1981 ataaattccg cccnnnnnnn nnnnctccct cccccccccc taacgttact ggccgaagcc
     2041 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     2101 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     2161 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     2221 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     2281 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     2341 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     2401 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     2461 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     2521 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     2581 acaaccatgg ttcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     2641 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     2701 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     2761 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     2821 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     2881 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     2941 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagtcgacga taaaataaaa
     3001 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     3061 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     3121 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     3181 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     3241 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     3301 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     3361 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     3421 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     3481 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     3541 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     3601 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     3661 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     3721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4321 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4621 tgcggccggc cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4681 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4741 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4801 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4861 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4921 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4981 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5041 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5101 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5161 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5221 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5281 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5341 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5401 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5461 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5521 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5581 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5641 cacatttccc cgaaaagtgc cacctgcata atgaaagacc ccacctgtag gtttggcaag
     5701 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagaaaag
     5761 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5821 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     5881 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5941 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6001 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     6061 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     6121 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     6181 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     6241 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
     6301 gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt
     6361 gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct
     6421 gtatctggcg gacccgtggt ggaactgacg agttcggaac acccggccgc aaccctggga
     6481 gacgtcccag ggacttcggg ggccgttttt gtggcccgac ctgagtccaa aaatcccgat
     6541 cgttttggac tctttggtgc acccccctaa taggagggat atgtggttct ggtaggagac
     6601 gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg ggaccgaagc
     6661 cgcgccgcgc gtcttgtctg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     6721 ctgtatttgt ctgaaaat
//
