LOCUS       sequence-297056-        8573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NDT1 in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8573)
  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 8573)
  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: YCplac33-NAT_NDT1
FEATURES             Location/Qualifiers
     source          1..8573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2203..2220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2203..2219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2212..2234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    2976..4138
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3301..4138
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     promoter        5078..5420
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      6000..6197
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6366..6384)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6452..6556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6557..6625,6626..7417)
                     /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     complement(6775..6794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7588..8176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8077..8096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8330..8347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8464..8485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8500..8505,8506..8523,8524..8530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8538..8554
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8543..8565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(8562..8573,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8562..8573,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccgcgtcac
       61 caacgcagag tccatgccga tgccttgctg ctcaagcaca ctgaaatacg cctctagttg
      121 tgtctcaata tctgtcttta gaaccatcaa ttccgacagt ttgaagctgt ctatttgact
      181 aattcttgat gttaggctcg gatctatctt aacattggcc aacaatttac ttaattcttc
      241 ttcctccatt gtagttcgtt cttattctaa ttcttgctcc tttttactct ggtgataact
      301 tttctatctc tcaatgctga acattttttt tccctttatt atccaatatt ccaaccgtta
      361 taaagtaagc cttatcgaac acaatccacg gcgcacgcaa gggacagcaa atttcgagaa
      421 ttaatgaata gccagtagta agatccgtat tgataaaggt catagatccc aacctttatg
      481 tgagccagcc cgaagggccc tttaaatgac tctcataata tggtccaata gcaccaattt
      541 ctaggggtta cccaccggcc tagtgcttaa tcgaattctg aaaagaatct ctactacaga
      601 ctgccaaccg taatagtagt gagatcaaca ctgaaggcag cagtctttag cacaatgaca
      661 cagactgata atcctgtccc caactgcggt ttactgcccg agcagcagta ttgctctgca
      721 gaccatgaag agccactgtt gttgcatgaa gaacaattga tattccctga tcattcctcc
      781 caactgtcct cagcagatat catcgagccc atcaagatga acagcagtac tgagtcaatt
      841 atagggacaa cgctgcgaaa gaaatgggta ccactatcct caactcagat cacagctctt
      901 tccggcgcat ttgctggatt cttatcaggt gtggcagtat gtcccctcga tgttgccaaa
      961 acgcgattgc aagcacaagg actacaaact aggttcgaga acccctacta tagggggata
     1021 atggggacat taagtactat agtaagagac gaaggcccgc ggggcctcta caaagggctg
     1081 gtaccgattg tcctgggcta cttcccaacc tggatgatat acttctccgt gtatgaattc
     1141 agcaaaaagt tctttcacgg catcttccca cagtttgatt ttgttgctca gtcatgtgct
     1201 gcaatcacgg caggcgctgc atctaccacc ttgaccaacc caatctgggt tgtgaagaca
     1261 agacttatgc tgcaatcaaa cctcggtgag caccccacac attacaaagg cactttcgat
     1321 gcattcagaa agctatttta tcaggaaggg tttaaagcat tatatgcggg gctggtcccc
     1381 tcattattag ggctatttca tgtggctatc catttcccta tatacgaaga tttgaaggta
     1441 agatttcact gctattctcg ggagaacaac accaactcca tcaacttgca acggttgatc
     1501 atggcatcgt ccgtctctaa gatgattgca tcagcagtaa catatccgca cgaaatttta
     1561 cgaaccagaa tgcaactgaa atcagatata ccagattcca ttcaacgacg tctgttcccc
     1621 ctcattaaag caacttatgc acaagaggga ctaaagggat tttattctgg atttactact
     1681 aacctagtac gaaccattcc ggcctcggca atcactctag tgtcctttga gtatttcaga
     1741 aaccgcctag aaaatattag cactatggta atttaaagcc ttaaaattca cacacgccac
     1801 acattgagtg tgctatgaat atgcctgcgc taatatttta ggtattacag cttgtaaata
     1861 taccaaataa ggtaagcctc tcttttccta atactaaaag gatttgcatc gaacagccca
     1921 ccaaatgctt tgcgcacaca ttatatacac ccacatgatt ctatagacaa aatatactta
     1981 tacattttca tagtcttgac aatattctac attggccgtt ttcttggcac tattttagct
     2041 tatccttacc aaaatagtct cttttttaaa tactattttt gaagcggaaa taggtgcgaa
     2101 caaatgaatg gcttcaattt agtgcgataa agacgtaaag cataaaccag tgaagatcat
     2161 agcactaaga aggcgcctgt actgttcctc gtgttggaat tcactggccg tcgttttaca
     2221 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2281 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2341 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2401 ttcacaccgc ataatcgctg ggccattctc atgaagaata tcttgaattt attgtcatat
     2461 tactagttgg tgtggaagtc ctaatatcgg tgatcaatat agtggttgac atgctggcta
     2521 gtcaacattg agccttttga tcatgcaaat atattacggt attttacaat caaatatcaa
     2581 acttaactat tgactttata acttatttag gtggtaacat tcttataaaa aagaaaaaaa
     2641 ttactgcaaa acagtactag cttttaactt gtatcctagg ttatctatgc tgtctcacca
     2701 tagagaatat tacctatttc agaatgtatg tccatgattc gccgggtaaa tacatataat
     2761 acacaaatct ggcttaataa agtctataat atatctcata aagaagtgct aaattggcta
     2821 gtgctatata tttttaagaa aatttctttt gactaagtcc atatcgactt tgtaaaagtt
     2881 cacattagca tacatatatt acacgagcca gaaatagtaa cttttgccta aatcacaaat
     2941 tgcaaaattt aattgcttgc aaaaggtcac atgcttataa tcaacttttt taaaaattta
     3001 aaatactttt ttatttttta tttttaaaca taaatgaaat aatttattta ttgtttatga
     3061 ttaccgaaac ataaaacctg ctcaagaaaa agaaactgtt ttgtccttgg aaaaaaagca
     3121 ctacctagga gcggccaaaa tgccgaggct ttcatagctt aaactcttta cagaaaatag
     3181 gcattataga tcagttcgag ttttcttatt cttccttccg gttttatcgt cacagtttta
     3241 cagtaaataa gtatcacctc ttagagttcg atgataagct gtcaaacatg agaattaatt
     3301 ccacatgtta aaatagtgaa ggagcatgtt cggcacacag tggaccgaac gtggggtaag
     3361 tgcactaggg tccggttaaa cggatctcgc attgatgagg caacgctaat tatcaacata
     3421 tagattgtta tctatctgca tgaacacgaa atctttactt gacgacttga ggctgatggt
     3481 gtttatgcaa agaaaccact gtgtttaata tgtgtcactg tttgatatta ctgtcagcgt
     3541 agaagataat agtaaaagcg gttaataagt gtatttgaga taagtgtgat aaagttttta
     3601 cagcgaaaag acgataaata caagaaaatg attacgagga tacggagaga ggtatgtaca
     3661 tgtgtattta tatactaagc tgccggcggt tgtttgcaag accgagaaaa ggctagcaag
     3721 aatcgggtca ttgtagcgta tgcgcctgtg aacattctct tcaacaagtt tgattccatt
     3781 gcggtgaaat ggtaaaagtc aaccccctgc gatgtatatt ttcctgtaca atcaatcaaa
     3841 aagccaaatg atttagcatt atctttacat cttgttattt tacagatttt atgtttagat
     3901 cttttatgct tgcttttcaa aaggcttgca ggcaagtgca caaacaatac ttaaataaat
     3961 actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt
     4021 cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc
     4081 taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagctct
     4141 cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg
     4201 tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac
     4261 tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga
     4321 actcttggta ttcttgccac gactcatctc catgcagttg gacgatcgat gataagctgt
     4381 caaacatgag aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     4441 agtatacatg catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa
     4501 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     4561 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     4621 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     4681 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     4741 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     4801 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     4861 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     4921 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     4981 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     5041 gcctttagcg gcttaactgt gccctcgccc ggccagcgac atggaggccc agaataccct
     5101 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
     5161 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
     5221 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
     5281 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
     5341 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
     5401 cacatccgaa cataaacaac tatgggcaca accttggatg atactgccta caggtacaga
     5461 acctctgtgc cgggagacgc tgaggcaata gaggcactag atggctcctt cacaacggac
     5521 accgttttta gagttactgc cacgggagat gggttcactt tacgtgaagt cccggttgac
     5581 cctccattga caaaagtctt tccagacgat gagtctgacg acgaatcaga cgacggggaa
     5641 gatggagatc cagactctag aacttttgtt gcttatggtg atgatggcga tttggccggg
     5701 ttcgtcgtcg tgagctactc tggttggaat aggaggttga cggtcgagga catcgaggtt
     5761 gcacctgagc atagaggaca tggggtgggc agagccttga tgggcctggc tacagaattt
     5821 gctcgtgaac gtggggcggg gcatctatgg cttgaagtca caaatgtcaa tgctcccgca
     5881 atacatgcat atagacgtat gggttttacc ctgtgcgggc ttgatacagc tttgtacgat
     5941 ggaaccgcta gtgacggtga acaagcctta tatatgtcca tgccttgccc ttaatcagta
     6001 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     6061 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     6121 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     6181 tgctggtcgc tatactgctg tcgattccat atgcgtatat ataccaatct aagtctgtgc
     6241 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     6301 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctaatt cttgaagacg
     6361 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6421 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6481 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6541 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6601 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6661 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6721 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6781 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6841 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6901 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6961 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7021 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7081 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7141 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7201 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7261 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7321 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7381 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7441 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7501 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7561 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7621 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7681 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7741 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7801 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7861 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7921 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7981 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8041 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8101 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8161 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8221 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8281 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     8341 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     8401 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     8461 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     8521 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gct
//
