LOCUS       sequence-320749-        7235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the C. auris MNase control strain, the URA3 auxotrophy marker of
            the CIp-pACT1-3xFLAG-MNase-SV40-CYC was replaced by SAT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7235)
  AUTHORS   Tebbji F, Khemiri I, Sellam A
  TITLE     High-Resolution Genome-Wide Occupancy in Candida spp. Using 
            ChEC-seq.
  JOURNAL   mSphere. 2020 Oct 14;5(5). pii: 5/5/e00646-20. doi: 
            10.1128/mSphere.00646-20.
  PUBMED    33055256
REFERENCE   2  (bases 1 to 7235)
  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: "mSphere."; 
            date: "2020-10-14"; volume: "5(5). pii: 5/5/e00646-20. doi"; pages: 
            " 10.1128/mSphere.00646-20"
COMMENT     Sequence Label: Cip-pAct1-Mnase-SAT1
FEATURES             Location/Qualifiers
     source          1..7235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(746..765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(747..765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(772..789)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(772..788)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(781..803)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(929..1384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1066..1087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1278..1297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1411..1515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1516..1584,1585..2376)
                     /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(1734..1753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2547..3135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3036..3055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3289..3306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3423..3444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3459..3464,3465..3482,3483..3489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3497..3513
                     /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     3502..3524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3521..3537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3521..3537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3558..3576
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             4640..4705
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4712..5158
                     /codon_start=1
                     /product="micrococcal nuclease from Staphylococcus aureus"
                     /label=MNase
                     /translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
                     HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
                     RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     CDS             5162..5182
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctccccaaca tcagcactcc tattcaatcc acctcatcgc agctatttcc aaggcaacgg
       61 aaccacatca cgcaagctcc agcgccccac aagcgctgcc tggcggaagc tacaagaatt
      121 acggtttttt tttttttttt tttttttgga atggccatga cgaaagctac aaggcaatga
      181 attggaccaa ttggaccaat tgaagggttt ttgagaattt ttgagcaatt tctctttgtt
      241 ctgttcacaa cttttctgac ttctacgttg aacaaggcct acggagagta gtgtgccgag
      301 tgggcgcagg tttgcaacct cacaccactt cgccccacac tgcgctcaag cgccaaagcg
      361 gccagcagaa aaaaaaaaag gcgttgaaca gggctgaagt cgggttaggg ttggcagaat
      421 tggtccagta tatttttgac tttgcttaag tatgggagtt ttagagtaga gggtgggata
      481 ggggggaggg gggagttccg agggcaggca aatgggcact tggggggagt tatcggcgac
      541 ttccgagggt ggcagcttct tctcgctgac ttccttcgtt cgaaggagtg agagccctcg
      601 gatttcccgc gctcagtcag tgggcccatg ggtgggttcg atattgcaac tagggaggcc
      661 cccctcggcg agtcacgtga tccgttctca cgacattgtc gcgggctgct gcccccggca
      721 cctcccgcgg tggagctcca attcgcccta tagtgagtcg tattacaatt cactggccgt
      781 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      841 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      901 acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc
      961 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1021 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1081 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1141 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1201 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1261 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1321 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1381 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1501 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1561 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1621 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1681 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1741 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1801 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1861 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1921 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1981 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2041 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2101 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2161 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2221 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2281 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2341 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3241 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3301 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3361 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3421 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     3481 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     3541 gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt accgggcccc
     3601 ccctcgagct attaagatca ccagcctcgt ttataataaa cttagtctaa tatttctttt
     3661 ggaccaaaat agaaactgtt ctcaggtggc aaaaaacagc ttaaagtctg tgttaaaaaa
     3721 agatctcggc tatgccaatc aaaaaggagt gacattatta tgtaaaagaa gaaatgttgt
     3781 aattccgcga aacaacaatt ttaatgctta ctgtgccgcc accaatatcc tatggccaag
     3841 gggcctttaa acatgccacg gaaatataca gtcaaagtaa acctactatg tctttagagc
     3901 cttcaggatc aattgttttt aaatgaatga ctgcaaaact agaaaagtag cctcttacat
     3961 aagcaagtca tttgtacacc gagagtttag taaccagtga agcaagtata tctggcaaac
     4021 ctgaaactta tctgtgaatt ataaagctca taagtaaaaa gtgtagaaaa gaagataagg
     4081 cagaccaaat acaaatttgc aacagtacaa cgaatatttc agcgtcaaaa ctagagaata
     4141 ataaagaaaa cgatcttttc aaaaagaaaa aaccttttag ttttcctttg ttgttgttgt
     4201 gggtgtgtgc tatttatatt atatagttta ctcataatac cataaaatat tcggtttgat
     4261 taggttattt taataagcta atttgtttct aatcgtgtaa tttatgctgt gtatattaag
     4321 tagtgtgtgc actgcccaaa aatgtttgtt gtttatagtc ggttaaagag aaaaaagaaa
     4381 aaaagatcca tacacacacg ttaattagtt gttcaacgta atacactcat attttgttct
     4441 tatttgcttt cggtcgctgt tctcaccaag atttattgcc aacgaaacaa ttttttttta
     4501 tatattttca gatttttctt tttttccttt cctttccttt tctaattttc actcctggtt
     4561 ttctttcttt cttagaaaca ttatctcgat attaatatta aaaaaatata atcattcaaa
     4621 aagcttgact acgcgtatgg actacaaaga ccacgacgga gactataagg atcacgatat
     4681 tgactataag gacgatgatg ataaagccgc cgccacctca acaaaaaaac ttcacaaaga
     4741 acctgcaacc ttaataaagg caattgacgg ggacactgtc aaattgatgt ataaaggtca
     4801 gccaatgacc ttccgtttac ttttagtcga tactcctgag actaagcatc ctaagaaagg
     4861 tgtagagaaa tatggtcctg aagccagtgc tttcacaaag aaaatggttg agaatgcaaa
     4921 gaaaattgag gtagagttcg acaaaggaca aagaaccgac aagtatggta gagggttggc
     4981 ctacatctac gcagatggaa agatggttaa cgaagcatta gtccgacagg gtttggctaa
     5041 agttgcctat gtgtataagc ctaacaatac ccacgaacag caccttcgta agtcagaagc
     5101 tcaagctaaa aaagaaaagt tgaatatctg gtcagaggat aatgcagact ctggtcaacc
     5161 acctaagaaa aaacgtaaag tataggctag caggaccacc tttgattgta aatagtaata
     5221 attaccaccc ttatctaatt atttatttaa cttatttatt tatttattat acatatatac
     5281 aaatctaata aagtgaaaat ctcccccttc acacttcaca tatgttaggc gtcatcctgt
     5341 gctcccgaga accagtacca gtacatcgct gtttcgttcg agacttgagg tctagtttta
     5401 tacgtgaaga ggtcaatgcc gccgagagta aagccacatt ttgcgtacaa attgcaggca
     5461 ggtacattgt tcgtttgtgt ctctaatcgt atgccaagga gctgtctgct tagtgcccac
     5521 tttttcgcaa attcgatgag actgtgcgcg actcctttgc ctcggtgcgt gtgcgacaca
     5581 acaatgtgtt cgatagaggc tagatcgttc catgttgagt tgagttcaat cttcccgaca
     5641 agctcttggt cgatgaatgc gccatagcaa gcagagtctt catcagagtc atcatccgag
     5701 atgtaatcct tccggtaggg gctcacactt ctggtagata gttcaaagcc ttggtcggat
     5761 aggtgcacat cgaacacttc acgaacaatg aaatggttct cagcatccaa tgtttccgcc
     5821 acctgctcag ggatcaccga aattttcata tgagaaccgt tatcgataac taaagcagca
     5881 acttcttcta taaaaatggg ttagtatgac agtcatttaa ataaggaatt tttcagttgg
     5941 cttggtttca attcaatgtt cgtttttttt ttttcttgct gtgtttgtgt ttgtgttgtt
     6001 tatagttgtg tgcactgagc gtcgaaaaaa aaaattcata gtgagccggg aaatctgtat
     6061 agcccagata acaacacaag tccaaactag aaactcgtca aacaccaaaa gcaatgttga
     6121 atcaattgcc ttgcacaagt acacgtagga aaacataaaa cattgcaatt ttgaatattg
     6181 agccttttgt cgtaacattg attgatagga ttactcaccg aatggttttg aaaccactgc
     6241 cgacagatca atcaatcaat caaaaaacgt gaactttgaa aaaggggaag aacagataca
     6301 ttgaagttag ccatttccac tgatcgtcac aacatatctg ataaattact ttcaaaatta
     6361 taagctgatg tgtgtgtatt attaatgtga cagtaacatc ccaaacgaga aatattatgt
     6421 cgacaacaaa aaagtttgat ctgaattgaa aatgaagttt tcccacccta cccatttgtc
     6481 atattgaaac caatcaactg attaatcaat caattagaat tgaagctaaa ctaaaacata
     6541 ccaccgtcca ttttgaatga ttatattttt ttaatattaa tatcgagata atgtttctaa
     6601 gaaagaaaga aaaccaggag tgaaaattag aaaaggaaag gaaaggaaaa aaagaaaaat
     6661 ctgaaaatat ataaaaaaaa attgtttcgt tggcaataaa tcttggtgag aacagcgacc
     6721 gaaagcaaat aagaacaaaa tatgagtgta ttacgttgaa caactaatta acgtgtgtgt
     6781 atggatcttt ttttcttttt tctctttaac cgactataaa caacaaacat ttttgggcag
     6841 tgcacacact acttaatata cacagcataa attacacgat tagaaacaaa ttagcttatt
     6901 aaaataacct aatcaaaccg aatattttat ggtattatga gtaaactata taatataaat
     6961 agcacacacc cacaacaaca acaaaggaaa actaaaaggt tttttctttt tgaaaagatc
     7021 gttttcttta ttattctcta gttttgacgc tcgacgcggc cgctctaact accgctcttc
     7081 caagcttcgc ttcctcgctt cggcaatttc aaccaaccta gttcacttac atttacgact
     7141 cttcggcctg cctttctctc tccactgggc caaacttgcc ctaactccca cgccagcgcc
     7201 agcaccgcca gcaattgccc ttttggcggt tttct
//
