LOCUS       Exported                6445 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression in S. cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    YCplac33-NAT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6445)
  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   Science Advances  09 Sep 2020: eabe5310
REFERENCE   2  (bases 1 to 6445)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6445
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             216..521
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIIAGPFS"
     misc_feature    233..289
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(290..307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    1063..2225
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      1388..2225
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     promoter        3165..3507
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4087..4284
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4453..4471)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4539..4643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4644..5504
                     /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(4862..4881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5675..6263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6164..6183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6417..6434
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
      481 gcggtatttc acaccgcata atcgctgggc cattctcatg aagaatatct tgaatttatt
      541 gtcatattac tagttggtgt ggaagtccta atatcggtga tcaatatagt ggttgacatg
      601 ctggctagtc aacattgagc cttttgatca tgcaaatata ttacggtatt ttacaatcaa
      661 atatcaaact taactattga ctttataact tatttaggtg gtaacattct tataaaaaag
      721 aaaaaaatta ctgcaaaaca gtactagctt ttaacttgta tcctaggtta tctatgctgt
      781 ctcaccatag agaatattac ctatttcaga atgtatgtcc atgattcgcc gggtaaatac
      841 atataataca caaatctggc ttaataaagt ctataatata tctcataaag aagtgctaaa
      901 ttggctagtg ctatatattt ttaagaaaat ttcttttgac taagtccata tcgactttgt
      961 aaaagttcac attagcatac atatattaca cgagccagaa atagtaactt ttgcctaaat
     1021 cacaaattgc aaaatttaat tgcttgcaaa aggtcacatg cttataatca acttttttaa
     1081 aaatttaaaa tactttttta ttttttattt ttaaacataa atgaaataat ttatttattg
     1141 tttatgatta ccgaaacata aaacctgctc aagaaaaaga aactgttttg tccttggaaa
     1201 aaaagcacta cctaggagcg gccaaaatgc cgaggctttc atagcttaaa ctctttacag
     1261 aaaataggca ttatagatca gttcgagttt tcttattctt ccttccggtt ttatcgtcac
     1321 agttttacag taaataagta tcacctctta gagttcgatg ataagctgtc aaacatgaga
     1381 attaattcca catgttaaaa tagtgaagga gcatgttcgg cacacagtgg accgaacgtg
     1441 gggtaagtgc actagggtcc ggttaaacgg atctcgcatt gatgaggcaa cgctaattat
     1501 caacatatag attgttatct atctgcatga acacgaaatc tttacttgac gacttgaggc
     1561 tgatggtgtt tatgcaaaga aaccactgtg tttaatatgt gtcactgttt gatattactg
     1621 tcagcgtaga agataatagt aaaagcggtt aataagtgta tttgagataa gtgtgataaa
     1681 gtttttacag cgaaaagacg ataaatacaa gaaaatgatt acgaggatac ggagagaggt
     1741 atgtacatgt gtatttatat actaagctgc cggcggttgt ttgcaagacc gagaaaaggc
     1801 tagcaagaat cgggtcattg tagcgtatgc gcctgtgaac attctcttca acaagtttga
     1861 ttccattgcg gtgaaatggt aaaagtcaac cccctgcgat gtatattttc ctgtacaatc
     1921 aatcaaaaag ccaaatgatt tagcattatc tttacatctt gttattttac agattttatg
     1981 tttagatctt ttatgcttgc ttttcaaaag gcttgcaggc aagtgcacaa acaatactta
     2041 aataaatact actcagtaat aacctatttc ttagcatttt tgacgaaatt tgctattttg
     2101 ttagagtctt ttacaccatt tgtctccaca cctccgctta catcaacacc aataacgcca
     2161 tttaatctaa gcgcatcacc aacattttct ggcgtcagtc caccagctaa cataaaatgt
     2221 aagctctcgg ggctctcttg ccttccaacc cagtcagaaa tcgagttcca atccaaaagt
     2281 tcacctgtcc cacctgcttc tgaatcaaac aagggaataa acgaatgagg tttctgtgaa
     2341 gctgcactga gtagtatgtt gcagtctttt ggaaatacga gtcttttaat aactggcaaa
     2401 ccgaggaact cttggtattc ttgccacgac tcatctccat gcagttggac gatcgatgat
     2461 aagctgtcaa acatgagaat tgggtaataa ctgatataat taaattgaag ctctaatttg
     2521 tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct
     2581 tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct
     2641 tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc
     2701 atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg
     2761 tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa
     2821 gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc
     2881 agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt
     2941 tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg
     3001 tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt
     3061 gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc
     3121 ggataatgcc tttagcggct taactgtgcc ctcgcccggc cagcgacatg gaggcccaga
     3181 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     3241 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     3301 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     3361 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     3421 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     3481 ctcacatcac atccgaacat aaacaactat gggcacaacc ttggatgata ctgcctacag
     3541 gtacagaacc tctgtgccgg gagacgctga ggcaatagag gcactagatg gctccttcac
     3601 aacggacacc gtttttagag ttactgccac gggagatggg ttcactttac gtgaagtccc
     3661 ggttgaccct ccattgacaa aagtctttcc agacgatgag tctgacgacg aatcagacga
     3721 cggggaagat ggagatccag actctagaac ttttgttgct tatggtgatg atggcgattt
     3781 ggccgggttc gtcgtcgtga gctactctgg ttggaatagg aggttgacgg tcgaggacat
     3841 cgaggttgca cctgagcata gaggacatgg ggtgggcaga gccttgatgg gcctggctac
     3901 agaatttgct cgtgaacgtg gggcggggca tctatggctt gaagtcacaa atgtcaatgc
     3961 tcccgcaata catgcatata gacgtatggg ttttaccctg tgcgggcttg atacagcttt
     4021 gtacgatgga accgctagtg acggtgaaca agccttatat atgtccatgc cttgccctta
     4081 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     4141 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     4201 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     4261 atgtgaatgc tggtcgctat actgctgtcg attccatatg cgtatatata ccaatctaag
     4321 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     4381 aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa gctaattctt
     4441 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4501 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4621 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4741 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4861 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4921 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5041 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5161 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5221 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5341 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5521 actcatatat actttagatt gatttaaaac ttcattttca atttaaaagg atctaggtga
     5581 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5641 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5701 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5761 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5821 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5881 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5941 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6061 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6121 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6181 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6241 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6301 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6361 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6421 agtcagtgag cgaggaagcg gaaga
//
