LOCUS       sequence-458318-        5272 bp    DNA     circular SYN 15-APR-2025
DEFINITION  B. dendrobatidis plasmid for integration of target sequence at URA3 
            locus, blasticidin S selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5272)
  TITLE     Homology-mediated stable transformation of Batrachochytrium 
            dendrobatidis illuminates chytrid cell biology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5272)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSB02
FEATURES             Location/Qualifiers
     source          1..5272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1327..1725
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(3108..3124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3108..3124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3121..3143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3296..3884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3376..3395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(4001..4025)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(4125..4916,4917..4985))
                     /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     4748..4767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4986..5090)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5189..5211
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5203..5220
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5204..5220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggcgattgg ttcgctgaac agatcttcat tgagcccaca tctgttggtg aggttgataa
       61 agtgctgcaa gattactcta gcactattag tagatctcgt cagacagatt tcagcacatc
      121 aacactcacc ggatctatgc ttttggctgt ggtatcagga aagctaagcg aaggaatcaa
      181 ctttgctgat gatatgggac ggtaagtggt tgtatttatt gcattagtac agtctgcaaa
      241 tgtttgttga tttctgggtt tatttatttc aatatatgca ctatatatgg atacatagag
      301 cagtgattat ggttggaatt ccatttgcca atattgcttc aatagaactg caagaacgga
      361 tgcgcttttt tgatgtgcag gtaaatgttt tgtacaaagt gtagtttgta ttgctttgat
      421 tggagatatt attgatttgc atgatttaac acgcataggc aaaagaaact ggatcgggca
      481 ttactggtaa agagtactac gaaaacattt gtatgcgtgc agtcaatcag tctgtaggca
      541 agtttatatg atgtggtact atacttgact atttaataac attgcaggcg tattgtgctt
      601 tgtaacaggt cgcgcaatca ggcacaagaa tgactatgca gtcattatat ttctagacat
      661 gcggttcaag cgagaccgta ttgtagctaa attgccggaa tggatccgta attgtggttt
      721 gacattcccg aatcagtttg ggccatctat tgccatgatt agcaaggtat gagccatatt
      781 tttgactgcg tttatcagat gtgtttggta ttctgactat tttattctcc atcactagtt
      841 tttcaattca catagaaagc ccaccaacaa aaaagaatag ccatttgaat ttaccattgt
      901 attcatgtaa aacgaaataa ttggtttgga atggccgagt attcgttgta ctttcaattc
      961 tttcaaacaa cgactatttt ataatgcaaa ctataagttt aaactggatt gtttggtaca
     1021 attctacact gcacaatcac ccaattcaac tacaatccaa atgttgggtt caactgccat
     1081 caaactcctg tattctaata tcagatacgt tatagttcaa gaaaacaaag aatgcaactg
     1141 ttggtttgat atcaatcact aaaagaacag aacttggatt agtaaatttg gtaaaaatta
     1201 tcatttcgtg agaccgattt attctatgat tttttttcca ctttacgtga atacgacaac
     1261 acattagcgc caattgagcc gattgaaggt cttgaacact tgctgataat ttaaactaaa
     1321 accggtatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
     1381 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
     1441 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
     1501 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
     1561 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
     1621 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
     1681 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctgactcga ggttggctca
     1741 tccactacac gtacaatcta aataaatagc tatttgctaa tctgtgtatt tccaagactt
     1801 tattctcaca tcagtactct accaatgtgc tattagagtt aattatttgc taaaagcgat
     1861 cctagctctg cgctgcgttc ggtagcacgc tggacggctt cagcaaagat ggctcgaatc
     1921 ttgttccctt cgagtgagtt gagtgctgcc tcggtggtac catttggaga tgtgactttg
     1981 cgtcttaatt cagctggatc ctcgcctgta tctgtcaaca tgcgacccgc tcctaaacaa
     2041 gtctgtgctg caagtccacg agcaacatct gcaggaagcc ccaatttcac tcctgcggct
     2101 tcgaggcact cgaccaacag gaaaaaataa gctggaccag atcctaacat gggtcaaaaa
     2161 agcatcagtt gatcaatcac caaatatatt ttacacaaaa tcagtgttca atacattgaa
     2221 atgaacttac cagatatgcc tgttacgaca tccaacagag attcctcctt gacccagtag
     2281 ttttgcttcg atacagactt gagcacagca gaaacaattt ccttttgatt ggtggtgact
     2341 gaaggagctg catacatacc cgttgctcct tcgacaacca acgcaggagt gtttggcatg
     2401 catcgcacaa ttgctggtga ctcggtcatc cctttcttgg ctgaacatgt caaccacttc
     2461 aacaagtttg aaatggtaat tccagcagca attgacacaa ttaatggctt gtgagtcaag
     2521 acggcgtccg aaatgccttc agcaacggat ctcatgactt gaggcttgac agcaagaata
     2581 agaacatcga caggagtcgt ttggctttga ttggtaaacg caattgcctg agtgtttgag
     2641 gtggtgacct gaactttaaa ccgtgaaact agacgagcac gagcttcgtc ccagggttct
     2701 gacacgataa tattggatgc ggaaagaccg gatttcaaca gtccgccaat aattgcacca
     2761 gccatattgc ctccaccaat aaagccaaga cgcaacccag taaagtctgt catgttgtaa
     2821 tgccaagact attgcgagaa ttgttgagca tcaaactaaa aataaaaaga agaatcaagc
     2881 caaccctcag gcagtccctt ccatccacgt gtcatgcaat ccattaccgt tcgaacatgc
     2941 agagctttct gctttattac acattggccg ctgtattcac gcaaagcaaa ctacattgat
     3001 ctcgatgact gttgattttg ctttaaagca atcgttgaat gctcgtgtgc taggctaggt
     3061 ggaggctcag tgatgataag tctgcgatgg tggatgcatg tgtcatggtc atagctgttt
     3121 cctgtgtgaa attgttatcc gctcagaggg cacaatccta ttccgcgcta tccgacaatc
     3181 tccaagacat taggtggagt tcagttcggc gtatggcata tgtcgctgga aagaacatgt
     3241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3721 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3901 tttctacggg gtctgacgct ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat
     3961 gggtaggggg cttcaaatcg tcctcgtgat accaattcgg agcctgcttt tttgtacaaa
     4021 cttgttgata atggcaattc aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4081 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4141 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4201 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4261 ccgcgagagc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4321 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4381 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4441 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4501 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4561 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4621 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4681 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4741 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4801 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4861 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4921 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4981 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5041 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5101 cgaaaagtgc cagatacctg aaacaaaacc catcgtacgg ccaaggaagt ctccaataac
     5161 tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     5221 catgcataat ccgcacgcat ctggaataag gaagtgccat tccgcctgac ct
//
