LOCUS       sequence-439432-        6611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector carrying a Candida auris centromere (CEN 4) with 
            Nourseothricin selectable marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6611)
  AUTHORS   Determann B 2nd, Fu J, Wickes BL
  TITLE     Development of a Shuttle Vector That Transforms at High Frequency 
            for the Emerging Human Fungal Pathogen: Candida auris.
  JOURNAL   J Fungi (Basel). 2024 Jul 11;10(7):477. doi: 10.3390/jof10070477.
  PUBMED    39057362
REFERENCE   2  (bases 1 to 6611)
  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: "J Fungi 
            (Basel)."; date: "2024-07-11"; volume: "10(7):477. doi"; pages: " 
            10.3390/jof10070477"
COMMENT     Sequence Label: p1153
FEATURES             Location/Qualifiers
     source          1..6611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1431..1447
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2496..2512)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2537..2556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2538..2556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2566..2583)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2566..2582)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2575..2597)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2724..3179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3021..3042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3205..3309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3310..3378,3379..4170)
                     /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(3528..3547)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4341..4929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4830..4849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5083..5100
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5217..5238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5253..5258,5259..5276,5277..5283)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5291..5307
                     /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     5296..5318
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5315..5331
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5315..5331
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5350..5370
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5352..5370
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atagtttgtg tatgcttctc ctatttttaa caaataatca atttttcaaa taatcaattt
       61 tttatcgatt ctattccggc ctaccatttt atcagaataa gatactattt gattaaaaaa
      121 actttgttcg aaggaaaatc tacttgaaca caggtatgat cggtcttaaa gttgcactaa
      181 tacgtagtcc gtcataaccg agtgacctgg aaaaagaaaa aaaaaaaaaa aaaaaaaaaa
      241 aatattaaag gaaaactagc aaagttacga tagtcgtcaa ttacagttcc tgctaaattt
      301 ataataaggt tatcttcgta ttcatttttt caagtcacgg tggatgcaca aagagcagct
      361 ccaacaccac taccgtcttt cgcaatctgc aaatgcaatt tcttgtttga tcccttcaac
      421 gggttaataa gattaaatgc atttaaaatc ttttcttgaa atcccgggta aaattcgact
      481 accgatccat cacagccaac ttcaaaggcg gccgctctag aactagtgcg ggtaccgcta
      541 tgccaaccaa aaaggagtga cattattatg taaaagaaga aatgttgtaa ttccgcgaaa
      601 caacaatttt aatgcttact gtgccgccac caatatccta tggccaaggg gcctttaaac
      661 atgccacgga aatatacagt caaagtaaac ctactatgtc tttagagcct tcaggatcaa
      721 ttgtttttaa atgaatgact gcaaaactag aaaagtagcc ccttacataa gcaagtcatt
      781 tgtacaccga gagtttagta accagtgaag caagtatatc tggcaaacct gaaacttatc
      841 tgtgaattat aaagctcata agtaaaaagt gtagaaaaga agataaggca gaccaaatac
      901 aaatttgcaa cagtacaacg aatatttcag cgtcaaaact agagaataat aaagaaaacg
      961 atcttttcaa aaagaaaaaa ccttttagtt ttcctttgtt gttgttgtgg gtgtgtgcta
     1021 tttatattat atagtttact cataatacca taaaatattc ggtttgatta ggttatttta
     1081 ataagctaat ttgtttctaa tcgtgtaatt tatgctgtgt atattaagta gtgtgtgcac
     1141 tgcccaaaaa tgtttgttgt ttatagtcgg ttaaagagaa aaaagaaaaa aagatccata
     1201 cacacacgtt aattagttgt tcaacgtaat acactcatat tttgttctta tttgctttcg
     1261 gtcgctgttc tcaccaagat ttattgccaa cgaaacaatt tttttttata tattttcaga
     1321 tttttctttt tttcctttcc tttccttttc taattttcac tcctggtttt ctttctttct
     1381 tagaaacatt atctcgatat taatattaaa aaaatataat cattcaaact cgaggtcgac
     1441 ggtatcgata agcttgatat cgaattcctg cagaattcag gaaggatcct acaattatgt
     1501 ctactacttt ggatgatact gcttatagat acagaacttc tgttccaggt gatgctgaag
     1561 ctattgaagc tttggatggt tctttcacta ccgatactgt ttttagagtt actgctactg
     1621 gtgatggttt cactttgaga gaagttccag ttgatccacc attgactaag gttttcccag
     1681 atgatgaatc cgatgatgaa tccgatgctg gtgaagatgg tgatccagat tctagaactt
     1741 tcgttgctta tggtgatgat ggtgatttgg ctggtttcgt tgttgtttct tattctggtt
     1801 ggaacagaag attgactgtt gaagatattg aagttgctcc agaacataga ggtcatggtg
     1861 ttggtagagc tttgatgggt ttggctactg aattcgccag agaaagaggt gctggtcatt
     1921 tgtggttgga agttaccaat gttaatgctc cagctattca tgcttataga agaatgggtt
     1981 tcactttgtg tggtttggat actgctttat acgatggtac tgcttccgat ggtgaacaag
     2041 ctttgtatat gtctatgcca tgtccataat aactcggggg gatccgagtg aaattctgga
     2101 aatctggaaa tctggttttg tattcttgtt attcttcttt ttgttattac atatataact
     2161 tgttactttt ttaaaaaaat ctttgtttat tttataaata tataaaacta aatttaagaa
     2221 aaagagaaaa atgttttatt tgagagattg aaattttact tgaatttagc ttagctttta
     2281 taaagtatta ttatgtaaaa aaacaaaaca aatatacatt aaaaagttaa gactataaaa
     2341 tagccaccca aggcatttct atatcttgtt gttgttgttt tcatcttctg tatcagagga
     2401 acttatttta ttattttcgt cacgggtatt ttctcttgtt tgatgattca tcccattcat
     2461 tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     2521 cccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     2581 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2641 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2701 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3481 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3541 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3721 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3841 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4141 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4981 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5041 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5101 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5161 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5221 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5281 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5341 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggagct
     5401 agaagcacgg gttccaacat cggagcacat tcgctaaaga aaaatatact cgtagatcat
     5461 atggatcaaa agtattaaac cactcatccg ttctactact tgtgtactta actacataac
     5521 atcataaaat ggaggcaatg aaaactattc agtgaacaaa gcaacctcgc atacgaaccg
     5581 ctacccaggc acaactcctc cgtatatcat tgcattagac aggaattatg ttgtatgtac
     5641 ttggattaaa aaataataaa atcgagttca aggtttgcgc attcagccga ttgactcgct
     5701 acataggcgt aatgaaagaa aaaaaaaaaa aaaaaattaa ttaaaagact actgcgaaaa
     5761 aaacgagtat gcaacatttc aaatgattta agcaaatgaa accgtcaagg ggtcactaac
     5821 ttaaattaaa tatgaaaaag tcacgaattc atttcttaca atacaagaat gaaaagatca
     5881 tattttgcct gaatcttttt tcttaaaaca tttggataca ttgatgtcgt ctatcgcttc
     5941 atacactggt agttttctcg cttatttttt acagatctat cgaacgggat aatttgctta
     6001 caccaaaata tgattgaagc aacgaatggt ggccatattt tgcttaagtg acatacgtta
     6061 tgccttcaag gacaatagat taataatcaa acacttgtat tctttacttt tctttctttg
     6121 gcggtgaagt atgaaacttt gtgcatgtgg atatgaatgt gtggagatac caacaatctt
     6181 ctcaaatatt tagaaatcaa ttcatgttgg tatcttcggt aaccaacaat cttcctctat
     6241 tccaactttt ctttgcaaac tcttttcaat cgcggtaaaa acgatcgagc gtatgccttt
     6301 ttggagacac acattcgaac ttcgtggtat ctaatctcac cgtcactcaa gccccgttaa
     6361 cttaaaaatc caacttcttt tgtttgcttt tcttagtgga ttttttctgt ctcctgataa
     6421 ttatgaatag aatatagagt cttccgggag aaggcaagac cattggaaga ctcgaaatta
     6481 ccttttgaag tgccgtagac ttccctgtcc attggcggct cgagacagca tgatacacca
     6541 gtctgcagaa atttgcttta tagaagtaga gtgcttatca acatatgaga tgaacataag
     6601 cttactttcg t
//
