LOCUS       sequence-194950-        6608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing the two fragments comprising the 550 bp NEUT5L 
            region with NAT1 marker. Integration by disruption of the region 
            targeted for recombination..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6608)
  AUTHORS   Gerami-Nejad M, Zacchi LF, McClellan M, Matter K, Berman J
  TITLE     Shuttle vectors for facile gap repair cloning and integration into a
            neutral locus in Candida albicans.
  JOURNAL   Microbiology. 2013 Mar;159(Pt 3):565-79. doi: 
            10.1099/mic.0.064097-0. Epub 2013 Jan 10.
  PUBMED    23306673
REFERENCE   2  (bases 1 to 6608)
  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: 
            "Microbiology."; date: "2013-03"; volume: "159(Pt 3):565-79. doi"; 
            pages: " 10.1099/mic.0.064097-0. Epub 2013 Jan 10"
COMMENT     Sequence Label: pDIS3
FEATURES             Location/Qualifiers
     source          1..6608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        191..471
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             472..1146
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(1386..1407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1913..1932
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2145..2167
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2159..2176
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2160..2176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2183..2202
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2183..2201
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2274..2471)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(3072..3415)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        complement(3484..3502)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3523..3539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3523..3539)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3536..3558)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3547..3563
                     /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     3581..3597
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3581..3597
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3631..3647)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3632..3648)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(join(3922..3928,3929..3946,3947..3952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3967..3988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4105..4122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4276..4864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4356..4375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5035..5826,5827..5895))
                     /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     5658..5677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5896..6000)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6037..6540
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6581..6599
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
      241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
      301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
      361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
      421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
      481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
      541 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
      601 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
      661 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
      721 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
      781 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
      841 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
      901 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
      961 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     1021 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     1081 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     1141 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
     1201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     1261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1561 cttgacgggg aaagcggccg gctaggcctt aacccactga attctacatc gaacaagaga
     1621 aacctttttt tttacttttt tcaaacttga acaaagaaaa gaaaaagaaa aagaaaaagc
     1681 aatcacagct tattaacgat tcgtgtttaa ataatcaatt gaaaggaaaa tagaagattt
     1741 ttgaaaaaaa gttattggta tttattaact taatatatca gtaacatcaa gagaaagaaa
     1801 ctgaattata aacagagtat gtgaagcaat tgcaattgca attattagag atccagaaaa
     1861 ctgaattgtg cttgaatacc acttgtttac cggcgaacgt ggcgagaaag gaagggaaga
     1921 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1981 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     2041 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2101 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2161 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccgaatt
     2221 cgagctcgtt taaacgtcga cactggatgg cggcgttagt atcgaatcga cagcagtata
     2281 gcgaccagca ttcacatacg attgacgcat gatattactt tctgcgcact taacttcgca
     2341 tctgggcaga tgatgtcgag gcgaaaaaaa atataaatca cgctaacatt tgattaaaat
     2401 agaacaacta caatataaaa aaactataca aatgacaagt tcttgaaaac aagaatcttt
     2461 ttattgtcag tactgactcg agttattatg gacatggcat agacatatac aaagcttgtt
     2521 caccatcgga agcagtacca tcgtataaag cagtatccaa accacacaaa gtgaaaccca
     2581 ttcttctata agcatgaata gctggagcat taacattggt aacttccaac cacaaatgac
     2641 cagcacctct ttctctggcg aattcagtag ccaaacccat caaagctcta ccaacaccat
     2701 gacctctatg ttctggagca acttcaatat cttcaacagt caatcttctg ttccaaccag
     2761 aataagaaac aacaacgaaa ccagccaaat caccatcatc accataagca acgaaagttc
     2821 tagaatctgg atcaccatct tcaccagcat cggattcatc atcggattca tcatctggga
     2881 aaaccttagt caatggtgga tcaactggaa cttctctcaa agtgaaacca tcaccagtag
     2941 cagtaactct aaaaacagta tcggtagtga aagaaccatc caaagcttca atagcttcag
     3001 catcacctgg aacagaagtt ctgtatctat aagcagtatc atccaaagta gtagacataa
     3061 ttgtaggatc cggttgttta tgttcggatg tgatgtgaga actgtatcct agcaagattt
     3121 taaaaggaag tatatgaaag aagaacctca gtggcaaatc ctaacctttt atatttctct
     3181 acaggggcgc ggcgtgggga caattcaacg cgtctgtgag gggagcgttt ccctgctcgc
     3241 aggtctgcag cgaggagccg taatttttgc ttcgcgccgt gcggccatca aaatgtatgg
     3301 acgcaaatga ttatacatgg ggatgtatgg gctaaatgta cgggcgacag tcacatcatg
     3361 cccctgagct gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg
     3421 ccgggtgacc cggcggggac gaggcaagct tgatatcccg cggtggagct ccagcttttg
     3481 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     3541 gtgaaattgt tatccgctca caattccaca tggcggccgc tctagaacta gtggatcccc
     3601 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggccgga
     3661 aggacgatga aggagagaaa gtagtaagac aatatgactt tggacagaca tgagataaaa
     3721 gttttaaagg acaagaaaag gttcgggtta aaagatgagt attgtatttt ttcaaaatat
     3781 caatgctttt tctgttcttg ggttagtggt accaccattg gtgataaaac ctttggggag
     3841 cctcctccga gctttctctt tgttactttt tttctcaact gatacttgtc attcttacta
     3901 caattacagg cctagccggc ccaacatagg agccggaagc ataaagtgta aagcctgggg
     3961 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4021 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4081 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4141 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4201 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4261 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4681 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5101 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5161 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5221 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5281 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5341 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5401 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5461 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5521 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5581 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5641 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5701 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5761 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5821 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5881 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5941 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6001 cacatttccc cgaaaagtgc cacctgggtc cttttcatca cgtgctataa aaataattat
     6061 aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa
     6121 agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat
     6181 actacctttt atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt
     6241 ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga
     6301 atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt
     6361 tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac tcttctacct
     6421 tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac acagagtaaa
     6481 ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag gcaagcgatc
     6541 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6601 ctttcgtc
//
