LOCUS       sequence-320752-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NheI-MluI-digested 3xFLAG-MNase-SV40 fragment was cloned into the 
            CIp-pACT1-CYC vector to ensure constitutive expression of MNase in 
            C. albicans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6804)
  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 6804)
  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
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     365..382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(535..540,541..558,559..565)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    573..589
                     /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     578..600
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     597..613
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     597..613
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        634..652
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1716..1781
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1788..2234
                     /codon_start=1
                     /product="micrococcal nuclease from Staphylococcus aureus"
                     /label=MNase
                     /translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
                     HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
                     RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     CDS             2238..2258
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3195..3211)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3195..3211)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4626..4645)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4627..4645)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4652..4669)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4652..4668)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4661..4683)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4809..5264)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4946..4967)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5158..5177
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5291..5395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5396..5464,5465..6256)
                     /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(5614..5633)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6427..6804,1..211)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
       61 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      541 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      601 aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa gggaacaaaa
      661 gctggtaccg ggccccccct cgagctatta agatcaccag cctcgtttat aataaactta
      721 gtctaatatt tcttttggac caaaatagaa actgttctca ggtggcaaaa aacagcttaa
      781 agtctgtgtt aaaaaaagat ctcggctatg ccaatcaaaa aggagtgaca ttattatgta
      841 aaagaagaaa tgttgtaatt ccgcgaaaca acaattttaa tgcttactgt gccgccacca
      901 atatcctatg gccaaggggc ctttaaacat gccacggaaa tatacagtca aagtaaacct
      961 actatgtctt tagagccttc aggatcaatt gtttttaaat gaatgactgc aaaactagaa
     1021 aagtagcctc ttacataagc aagtcatttg tacaccgaga gtttagtaac cagtgaagca
     1081 agtatatctg gcaaacctga aacttatctg tgaattataa agctcataag taaaaagtgt
     1141 agaaaagaag ataaggcaga ccaaatacaa atttgcaaca gtacaacgaa tatttcagcg
     1201 tcaaaactag agaataataa agaaaacgat cttttcaaaa agaaaaaacc ttttagtttt
     1261 cctttgttgt tgttgtgggt gtgtgctatt tatattatat agtttactca taataccata
     1321 aaatattcgg tttgattagg ttattttaat aagctaattt gtttctaatc gtgtaattta
     1381 tgctgtgtat attaagtagt gtgtgcactg cccaaaaatg tttgttgttt atagtcggtt
     1441 aaagagaaaa aagaaaaaaa gatccataca cacacgttaa ttagttgttc aacgtaatac
     1501 actcatattt tgttcttatt tgctttcggt cgctgttctc accaagattt attgccaacg
     1561 aaacaatttt tttttatata ttttcagatt tttctttttt tcctttcctt tccttttcta
     1621 attttcactc ctggttttct ttctttctta gaaacattat ctcgatatta atattaaaaa
     1681 aatataatca ttcaaaaagc ttgactacgc gtatggacta caaagaccac gacggagact
     1741 ataaggatca cgatattgac tataaggacg atgatgataa agccgccgcc acctcaacaa
     1801 aaaaacttca caaagaacct gcaaccttaa taaaggcaat tgacggggac actgtcaaat
     1861 tgatgtataa aggtcagcca atgaccttcc gtttactttt agtcgatact cctgagacta
     1921 agcatcctaa gaaaggtgta gagaaatatg gtcctgaagc cagtgctttc acaaagaaaa
     1981 tggttgagaa tgcaaagaaa attgaggtag agttcgacaa aggacaaaga accgacaagt
     2041 atggtagagg gttggcctac atctacgcag atggaaagat ggttaacgaa gcattagtcc
     2101 gacagggttt ggctaaagtt gcctatgtgt ataagcctaa caatacccac gaacagcacc
     2161 ttcgtaagtc agaagctcaa gctaaaaaag aaaagttgaa tatctggtca gaggataatg
     2221 cagactctgg tcaaccacct aagaaaaaac gtaaagtata ggctagcgtc cctatttatt
     2281 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattcgc gtagatccaa
     2341 ctcaagtaca acatggctgt cggtaaaaac aagagattgt ccaaaggaaa gaaaggatta
     2401 aaaaagaagg tcgttgaccc attcaccaga aaagattggt ttgacatcaa agctccaacc
     2461 acttttgaaa acagaaatgt tggtaaaact ttgatcaaca gatctaccgg tttaaagaat
     2521 gccgctgatg gcttgaaagg tagagttttc gaagtttgtt tggccgactt acaaggttcc
     2581 gaagaccact cttacagaaa aatcaaattg agagttgatg aagtccaagg taaaaacttg
     2641 ttgaccaact tccatgaggc ctccatggag gcctcatgga ggcctcatgg aggcctcatg
     2701 gaggcctcat ggtttggatt tcacttctga caaattaaga tcattggtca gaaaatggca
     2761 atcattagtt gaagctaatg tcactgttaa aacttccgac gattacgttt tgagagtttt
     2821 tgccattgct ttcaccaaaa gacaaccaaa ccaaatcaag aaaactactt acgctcaatc
     2881 ctctaaattg agagaagtca gaaagaagat gattgaaatt atgcaaagag aagtttccaa
     2941 ctgtacttta gctcaattaa cttccaaatt gattccagaa gtcattggcc gtgaaattga
     3001 aaaatccacc caaaccattt tcccattaca aaatgtccac atcagaaaag tcaaattgtt
     3061 gaaacaacca aaattcgact tgggttcatt attggctttg cacggtgaag gttcaaccga
     3121 agaaaaaggt aagaaagttt cttctggttt caaagatgtt gttttagaat ctgtttaagg
     3181 ataaggatct gggggatcca ctagttctag aaggaccacc tttgattgta aatagtaata
     3241 attaccaccc ttatttaatt atttatttaa cttatttatt tatttattat acatatatac
     3301 aaatctaata aagtgaaaat ctcccccttc acatttataa ttggccagtt tttttcaaat
     3361 aagcattcca accagcatct ctataccttt taccttcaat atctggatct cttcctttac
     3421 caaacaatcc tctaccaaca atgataatat cagttccagt gctaacaact tcatcaacag
     3481 ttctatattg ttgtcctaat ccatcacctt tatcatctaa tccaactcca ggtgtcataa
     3541 taagccaatc aaatccttct tcttgtccac ccatatcacg ttgggcaata aatccaataa
     3601 caaattcctt atcggattta gcaatttcaa cagttttttg agaatattct ccatatgcta
     3661 atgatcccac tgatgataat tcagctaaca tcaataaccc tcttggctct tggttggtgg
     3721 tggtttcttt agctccctgt tttaatcctt caactactcc attcccagtg acaccatgag
     3781 cattagtaat atctgcccaa ctactaattt tataaactcc accaatatat tgtttcttca
     3841 cggtattacc aatatcagca aattttctat cttcaaaaat cataaattga tgtttacgtg
     3901 aaagttctaa taatggttca atagtggatt cataggaaaa atcattgatt atatcaatat
     3961 gagtcttgat taagcataca taaggaccca atttatcaat taattcaagg aattccttag
     4021 tggtatcaac atcaattgat gcacataaat tggttttctt cagttccatt aatcgaaata
     4081 atcgttgtgc tactggtgag gcatgagttt ctgctctctc actataggtc ttagtgttga
     4141 ctgtcatatc ttttgtttgc tatttctatt tctatttctt tcttcaactg taaaaggggc
     4201 gatatataaa ccagtgtgta tggggttgtt gctcaggaat tgatcaatgt ttaaaagaat
     4261 ggttgaaatc atcattgaag aacaaaaaaa aagatttttt ctttcattat gttatctgta
     4321 taatatgcag gaaataagat tgcaaaaaaa tagccgtttt tttggatact atcaaacaag
     4381 aggaatgact aatacagaat tagtgttacg aatcaatggc actacagcaa ctttcaacaa
     4441 aataatctac gttcaaaata taatatatac tattaaataa aatcaaataa taaaataatt
     4501 ttaaagaaag tagtaccagc tctttttttt gtttccgttt ataccatcca aatcaattcc
     4561 tattagtcgg gatctaggcc tggccaagtc ggccgcggcc gccaccgcgg tggagctcca
     4621 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     4681 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     4741 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     4801 gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4861 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4921 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4981 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     5041 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5101 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5161 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5221 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     5281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5341 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5401 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5461 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5521 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5581 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5641 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5701 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5761 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5821 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5881 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5941 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6001 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6061 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6121 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6181 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6241 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6301 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6781 aacgacctac accgaactga gata
//
