LOCUS       sequence-261597-        6610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLAG-tagged GGC1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  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 6610)
  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-GGC1
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             41..457
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     387..409
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     425..441
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    467..841
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     779..795
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      870..879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             879..944
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(1909..2483,2484..2486,2487..2495)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2076..2093)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2303..2322
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2496..2891
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3062..3531
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3661..3678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3832..4420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3912..3931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4584..5375,5376..5444))
                     /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     5207..5226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5445..5549)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5579..6172
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6235..6592
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
       61 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
      121 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
      181 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
      241 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
      301 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
      361 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
      421 ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat atgagatctt
      481 atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa gagttactaa
      541 cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg aagtctggag
      601 acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc acccttaccg
      661 agtcggcgac acagtgtggg tccgccgaca ccagactaag aacctagaac ctcgctggaa
      721 aggaccttac acagtcctgc tgaccacccc caccgccctc aaagtagacg gcatcgcagc
      781 ttggatacac gccgcccacg tgaaggctgc cgaccccggg ggtggaccat cctctagact
      841 gccggatcta gctagttaat taaggatccg ccaccatgga ctacaaggac cacgacggtg
      901 actacaagga ccacgacatc gactacaagg acgacgacga caaggggcat ggcaccggca
      961 gcaccggcag cgaattcatg cctcataccg acaagaagca gtcaggcctc gcccggctct
     1021 tgggaagcgc aagtgctgga ataatggaga ttgccgtttt ccaccctgta gatacgatct
     1081 ctaagcggct tatgagtaat cacaccaaaa ttacgagtgg ccaggagctg aatagggtaa
     1141 tctttcgcga ccatttttct gaaccgctcg gaaagaggct gtttacactt ttccccggct
     1201 tgggttatgc cgcgtcttac aaggtgctgc agcgagtcta caaatatggg ggacagcctt
     1261 tcgcgaatga gttccttaat aaacactata aaaaagattt cgacaacctt tttggagaga
     1321 aaaccgggaa agccatgcga tccgccgccg cagggtcctt gatagggatt ggggaaatcg
     1381 ttttgctgcc tttggacgtg ctgaaaatta aaagacaaac gaatcccgag agttttaagg
     1441 gtcgggggtt catcaaaatt ctgcgggatg aaggcctttt caatctgtac cggggctggg
     1501 gctggactgc agcccggaat gctccggggt cctttgcctt gttcggcggc aatgcctttg
     1561 caaaagaata tatacttgga ttgaaggatt acagtcaagc aacgtggagt caaaacttta
     1621 tttcctccat agttggagcc tgcagctccc ttatcgtctc agctcccctg gatgtaatca
     1681 agacgcgaat ccagaaccga aactttgata accccgaatc cggcctccgc atagtcaaga
     1741 ataccctcaa gaatgaggga gtaacagcat tcttcaaggg tctcacacct aagttgctga
     1801 caacgggacc caagttggtt ttctctttcg cactcgctca atcactgatt ccgagattcg
     1861 ataacctgct gagtaaatga gcggccgcta cgtaaattcc gccccccccc cctctccctc
     1921 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1981 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     2041 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     2101 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     2161 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     2221 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     2281 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     2341 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     2401 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     2461 cctttgaaaa acacgatgat aatatggcca caaccatggt tcctttgtct caagaagaat
     2521 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     2581 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     2641 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     2701 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     2761 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     2821 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     2881 gggagggcta agtcgacgat aaaataaaag attttattta gtctccagaa aaagggggga
     2941 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     3001 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     3061 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3121 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     3181 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     3241 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     3301 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     3361 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     3421 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     3481 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca
     3541 tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca
     3601 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3661 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3721 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3781 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3841 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3901 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3961 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4021 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4081 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4141 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4201 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4261 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4321 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4381 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4441 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4501 atcaaaaagg atcttcacct agatcctttt gcggccggcc gcaaatcaat ctaaagtata
     4561 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4621 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4681 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4741 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4801 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4861 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4921 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4981 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5041 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5101 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5161 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5221 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5281 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5341 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5401 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5461 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5521 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgcataa
     5581 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     5641 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca gatggaacag
     5701 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     5761 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5821 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     5881 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     5941 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     6001 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gattgactga gtcgcccggg
     6061 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     6121 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
     6181 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
     6241 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
     6301 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
     6361 gttcggaaca cccggccgca accctgggag acgtcccagg gacttcgggg gccgtttttg
     6421 tggcccgacc tgagtccaaa aatcccgatc gttttggact ctttggtgca cccccctaat
     6481 aggagggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     6541 aatttttgct ttcggtttgg gaccgaagcc gcgccgcgcg tcttgtctgc tgcagcatcg
     6601 ttctgtgttg
//
