LOCUS       sequence-194948-        6607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector containing the two fragments comprising the 550 bp NEUT5L 
            region with NAT1 marker. Integration by duplication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6607)
  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 6607)
  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: pDUP3
FEATURES             Location/Qualifiers
     source          1..6607
                     /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(3921..3927,3928..3945,3946..3951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3966..3987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4104..4121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4275..4863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4355..4374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5034..5825,5826..5894))
                     /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     5657..5676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5895..5999)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6036..6539
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6580..6598
                     /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 aaacaagtgg tattcaagca caattcagtt
     1621 ttctggatct ctaataattg caattgcaat tgcttcacat actctgttta taattcagtt
     1681 tctttctctt gatgttactg atatattaag ttaataaata ccaataactt tttttcaaaa
     1741 atcttctatt ttcctttcaa ttgattattt aaacacgaat cgttaataag ctgtgattgc
     1801 tttttctttt tctttttctt ttctttgttc aagtttgaaa aaagtaaaaa aaaaggtttc
     1861 tcttgttcga tgtagaattc agtgggttac 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 atgcaaatga 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 gggggccgta
     3661 attgtagaag aatgacaagt atcagttgag aaaaaaagta acaaagagaa agctcggagg
     3721 aggctcccca aaggttttat caccaatggt ggtaccacta acccaagaac agaaaaagca
     3781 ttgatatttt gaaaaaatac aatactcatc ttttaacccg aaccttttct tgtcctttaa
     3841 aacttttatc tcatatctgc ccaaagtcat attgtcttac tactttctct ccttcatcgt
     3901 ccttccaggc ctagccggcc caacatagga gccggaagca taaagtgtaa agcctggggt
     3961 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4021 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4081 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5041 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5101 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5161 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5221 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5281 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5341 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5401 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5461 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5521 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5581 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5641 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5701 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5761 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5821 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5881 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5941 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6001 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata
     6061 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
     6121 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
     6181 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
     6241 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
     6301 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
     6361 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     6421 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     6481 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     6541 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6601 tttcgtc
//
