LOCUS       sequence-261683-        6541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged NMNAT1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6541)
  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 6541)
  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-NMNAT1
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..449
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     379..401
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     417..433
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    459..833
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     771..787
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      862..871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             871..936
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1855..2407
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1999..2016)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2226..2245
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2419..2814
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2985..3454
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3584..3601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3755..4343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3835..3854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4507..5298,5299..5367))
                     /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     5130..5149
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5368..5472)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5502..6095
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6158..6515
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
       61 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
      121 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
      181 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
      241 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
      301 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
      361 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
      421 atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg
      481 cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct
      541 ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg
      601 cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg
      661 acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt
      721 acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac
      781 acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctaga ctgccggatc
      841 tagctagtta attaaggatc cgccaccatg gactacaagg accacgacgg tgactacaag
      901 gaccacgaca tcgactacaa ggacgacgac gacaaggggc atggcaccgg cagcaccggc
      961 agcgaattca tggaaaattc cgagaagact gaagtggttc tccttgcttg tggttcattc
     1021 aatcccatca ccaacatgca cctcaggttg tttgagctgg ccaaggacta catgaatgga
     1081 acaggaaggt acacagttgt caaaggcatc atctctcctg ttggtgatgc ctacaagaag
     1141 aaaggactca ttcctgccta tcaccgggtc atcatggcag aacttgctac caagaactct
     1201 aaatgggtgg aagttgatac atgggaaagt cttcagaagg agtggaaaga gactctgaag
     1261 gtgctaagac accatcaaga gaaattggag gctagtgact gtgatcacca gcagaactca
     1321 cctactctag aaaggcctgg aaggaagagg aagtggactg aaacacaaga ttctagtcaa
     1381 aagaaatccc tagagccaaa aacaaaagct gtgccaaagg tcaagctgct gtgtggggca
     1441 gatttattgg agtcctttgc tgttcccaat ttgtggaaga gtgaagacat cacccaaatc
     1501 gtggccaact atgggctcat atgtgttact cgggctggaa atgatgctca gaagtttatc
     1561 tatgaatcgg atgtgctgtg gaaacaccgg agcaacattc acgtggtgaa tgaatggatc
     1621 gctaatgaca tctcatccac aaaaatccgg agagccctca gaaggggcca gagcattcgc
     1681 tacttggtac cagatcttgt ccaagaatac attgaaaagc ataatttgta cagctctgag
     1741 agtgaagaca ggaatgctgg ggtcatcctg gcccctttgc agagaaacac tgcagaagct
     1801 aagacatagg cggccgctac gtaaattccg cccccccccc ctcccccccc cctaacgtta
     1861 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     1921 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     1981 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2041 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2101 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2161 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2221 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2281 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     2341 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     2401 gataatatgg ccacaaccat ggttcctttg tctcaagaag aatccaccct cattgaaaga
     2461 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     2521 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     2581 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     2641 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     2701 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     2761 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagtcgac
     2821 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     2881 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     2941 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     3001 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     3061 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     3121 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     3181 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     3241 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     3301 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     3361 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     3421 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     3481 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     3541 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3601 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4321 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4441 cctagatcct tttgcggccg gccgcaaatc aatctaaagt atatatgagt aaacttggtc
     4501 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4561 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4621 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4681 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4741 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4801 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4861 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4921 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4981 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5041 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5101 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5161 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5221 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5281 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5341 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5401 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5461 aggggttccg cgcacatttc cccgaaaagt gccacctgca taatgaaaga ccccacctgt
     5521 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     5581 agaatagaaa agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     5641 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     5701 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     5761 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     5821 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     5881 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     5941 ctcactcggc gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     6001 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     6061 agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga
     6121 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt
     6181 gtctgtccga ttgtctagtg tctatgactg attttatgcg cctgcgtcgg tactagttag
     6241 ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttcgga acacccggcc
     6301 gcaaccctgg gagacgtccc agggacttcg ggggccgttt ttgtggcccg acctgagtcc
     6361 aaaaatcccg atcgttttgg actctttggt gcacccccct aataggaggg atatgtggtt
     6421 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     6481 tgggaccgaa gccgcgccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
     6541 c
//
