LOCUS       sequence-261724-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged NMNAT2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  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 6633)
  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-NMNAT2
FEATURES             Location/Qualifiers
     source          1..6633
                     /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(1980..2554,2555..2557,2558..2566)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2147..2164)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2374..2393
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2567..2962
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3133..3602
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3732..3749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3903..4491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3983..4002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4655..5446,5447..5515))
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5650..6243
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    join(6306..6633,1..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 cgagaccacc aagacccacg ttatcttgct cgcctgcggc agcttcaatc
     1081 ccatcaccaa agggcacatt cagatgtttg aaagagccag ggattatctg cacaaaactg
     1141 gaaggtttat tgtgattggc gggattgtct cccctgtcca cgactcctat ggaaaacagg
     1201 gcctcgtgtc aagccggcac cgtctcatca tgtgtcagct ggccgtccag aactctgatt
     1261 ggatcagggt ggacccttgg gagtgctacc aggacacctg gcagacgacc tgcagcgtgt
     1321 tggaacacca ccgggacctc atgaagaggg tgactggctg catcctctcc aatgtcaaca
     1381 caccttccat gacacctgtg atcggacagc cacaaaacga gaccccccag cccatttacc
     1441 agaacagcaa cgtggccacc aagcccactg cagccaagat cttggggaag gtgggagaaa
     1501 gcctcagccg gatctgctgt gtccgcccgc cggtggagcg tttcaccttt gtagatgaga
     1561 atgccaatct gggcacggtg atgcggtatg aagagattga gctacgcatc ctgctgctgt
     1621 gtggtagtga cctgctggag tccttctgca tcccagggct ctggaacgag gcagatatgg
     1681 aggtgattgt tggtgacttt gggattgtgg tggtgccccg ggatgcagcc gacacagacc
     1741 gaatcatgaa tcactcctca atactccgca aatacaaaaa caacatcatg gtggtgaagg
     1801 atgacatcaa ccatcccatg tctgttgtca gctcaaccaa gagcaggctg gccctgcagc
     1861 atggggacgg ccatgttgtg gattacctgt cccagccggt catcgactac atcctcaaaa
     1921 gccagctgta catcaatgcc tccggctagg cggccgctac gtaaattccg cccccccccc
     1981 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     2041 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     2101 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     2161 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     2221 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     2281 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     2341 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2401 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     2461 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2521 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg ttcctttgtc
     2581 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
     2641 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
     2701 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
     2761 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
     2821 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
     2881 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
     2941 tggttatgtg tgggagggct aagtcgacga taaaataaaa gattttattt agtctccaga
     3001 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     3061 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     3121 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3181 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     3241 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     3301 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     3361 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     3421 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     3481 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     3541 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     3601 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     3661 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3721 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccggc cgcaaatcaa
     4621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5641 cacctgcata atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     5701 ttgcaaggca tggaaaaata cataactgag aatagaaaag ttcagatcaa ggtcaggaac
     5761 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     5821 ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     5881 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca
     5941 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     6001 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     6061 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgattgactg
     6121 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     6181 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     6241 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg
     6301 gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc tatgactgat
     6361 tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg gacccgtggt
     6421 ggaactgacg agttcggaac acccggccgc aaccctggga gacgtcccag ggacttcggg
     6481 ggccgttttt gtggcccgac ctgagtccaa aaatcccgat cgttttggac tctttggtgc
     6541 acccccctaa taggagggat atgtggttct ggtaggagac gagaacctaa aacagttccc
     6601 gcctccgtct gaatttttgc tttcggtttg gga
//
