LOCUS       sequence-268961-        6406 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Integration of COX4-GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Marker for yeast 
            mitochondria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6406)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 6406)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM151
FEATURES             Location/Qualifiers
     source          1..6406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1038..1103
                     /codon_start=1
                     /product="mitochondrial presequence of yeast cytochfome c 
                     oxidase subunit IV (Hurt et al., 1984)"
                     /label=pre-CoxIV
                     /translation="MLSLRQSIRFFKPATRTLCSSR"
     CDS             1527..2243
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      2264..2451
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2319..2343)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2734..2767
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      3722..3919
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3973..4006
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4060..4079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4061..4079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4166..4183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4337..4925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4417..4436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5096..5887,5888..5956))
                     /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     5719..5738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5957..6061)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6129..6147
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6185..6207)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6307..6326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagcc tattggattg gcccaattgc
       61 tgccaaattt ggtgaatatg gtggtgatgt cgcaatgtgg ttaagtatgg cgttctcagg
      121 tgttgtatac cctccttgta gatacttaga gctacgtaaa tttggtcgtt aattgaaatg
      181 acataagtat atattaaaac tacggtgcag cggcaaactt tagtttagat aatattggca
      241 ggtagattca tggtatgtta aattacgctc agggaggata tttttttgta cacattatct
      301 tttcacatga aaatctaagc ctggtggaca attgaaatca tcgagtgata tactgacact
      361 ttgaacagta gcacaaatcg tttaatttgc tgtcacgatc taagttttct gtcagctgac
      421 tgtccaccaa taagatcagc ggattccaaa ggcgtccttc gaaaaagcga tatctcccat
      481 cttggttagt agcaaataaa gggaagagaa aataagacac actcaaggat aaaatacttc
      541 gcttcttcca agtataaaca caggaaagaa agatatacga agattagaat ttttttcata
      601 ctttttacaa aacatttttt tgtacttagg cacgctaatt gcgtaaaaat tagcataaac
      661 aggaaataag attattttga aatgtatgtt ataatcctct tattgaaaaa tacatgagat
      721 gtaggagaag aactaccagg aaagaagata tttaacagat cgaacgaaac ggaaaagaac
      781 taagagaaaa caaatctgga tttgacttta agaaaaaatt tatttgcaag tttaactcgt
      841 tccctgcttt tctatttggc ttgagacgat aattgcataa ttgtccaaat tgtccatcat
      901 ataatgaaga attcgatgcg aaatattccc tgtcactttc ctttagtcaa cgtattcttc
      961 tccctgaaga aacagtatac taacaatact cacccatttc gattttgatg ttgccataca
     1021 aatagataac aagcacaatg ctttcactac gtcaatctat aagatttttc aagccagcca
     1081 caagaacttt gtgtagctct agatatctgc ttcagcaaaa acccgtggtg aaaactgccc
     1141 aaaacttagc agaagttaat ggtccagaaa ctttgattgg tcctggtgct aaagagggta
     1201 ccgttccaac agacctagat caagaaactg gtttagctag gttagaatta ttgggtaaat
     1261 tagagggtat cgatgttttc gacaccaaac cattagattc gtccaggaag ggtaccatga
     1321 aagatccgat catcattgaa tcttatgatg attatcgtta tgtcggttgt acgggttctc
     1381 ccgctggttc acatactatt atgtggttaa aaccaactgt taacgaagtc gccagatgct
     1441 gggaatgtgg ttctgtttac aaactaaacc ctgttggtgt tccaaatgat gaccaccatc
     1501 acctttctct tgaagttgct gaagaaatgt ctaaaggtga agaattattc actggtgttg
     1561 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     1621 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
     1681 aattgccagt tccatggcca accttagtca ctactttaac ttatggtgtt caatgttttt
     1741 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     1801 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     1861 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     1921 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     1981 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
     2041 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     2101 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
     2161 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     2221 gtattgatga attgtacaaa taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     2281 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     2341 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     2401 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     2461 tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     2521 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     2581 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     2641 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcggacct
     2701 gcgagcaggg aaacgctccc ctcacagacg cgtataactt cgtataatgt atgctatacg
     2761 aagttattga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg
     2821 ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac
     2881 atcacatccg aacataaaca accatgacag tcaacactaa gacctatagt gagagagcag
     2941 aaactcatgc ctcaccagta gcacaacgat tatttcgatt aatggaactg aagaaaacca
     3001 atttatgtgc atcaattgat gttgatacca ctaaggaatt ccttgaatta attgataaat
     3061 tgggtcctta tgtatgctta atcaagactc atattgatat aatcaatgat ttttcctatg
     3121 aatccactat tgaaccatta ttagaacttt cacgtaaaca tcaatttatg atttttgaag
     3181 atagaaaatt tgctgatatt ggtaataccg tgaagaaaca atatattggt ggagtttata
     3241 aaattagtag ttgggcagat attactaatg ctcatggtgt cactgggaat ggagtagttg
     3301 aaggattaaa acagggagct aaagaaacca ccaccaacca agagccaaga gggttattga
     3361 tgttagctga attatcatca gtgggatcat tagcatatgg agaatattct caaaaaactg
     3421 ttgaaattgc taaatccgat aaggaatttg ttattggatt tattgcccaa cgtgatatgg
     3481 gtggacaaga agaaggattt gattggctta ttatgacacc tggagttgga ttagatgata
     3541 aaggtgatgg attaggacaa caatatagaa ctgttgatga agttgttagc actggaactg
     3601 atattatcat tgttggtaga ggattgtttg gtaaaggaag agatccagat attgaaggta
     3661 aaaggtatag agatgctggt tggaatgctt atttgaaaaa gactggccaa ttataatcag
     3721 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     3781 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     3841 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     3901 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tgtcgaaaac
     3961 gagctcgaat tcataacttc gtataatgta tgctatacga agttatatcg atgatatcag
     4021 atccactagt ggcctatgcg gccgcggatc tgccggtctc cctatagtga gtcgtattaa
     4081 tttcgataag ccaggttaac ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4141 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4741 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4981 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5041 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5101 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5161 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5221 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5281 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5341 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5401 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5461 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5521 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5581 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5641 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5701 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5761 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5821 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5881 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5941 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6001 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6061 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6121 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     6181 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6241 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6301 actatgcggc atcagagcag attgtactga gagtgcacca tatggacata ttgtcgttag
     6361 aacgcggcta caattaatac ataaccttat gtatcataca catacg
//
