LOCUS       sequence-468381-        4815 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid for deleting or C-terminally tagging endogenous genes with 
            mScarlet and selection on G418..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4815)
  AUTHORS   Colarusso A, Williams A, Gladfelter AS, Wirshing ACE, Lew DJ
  TITLE     Optimized vectors for genetic engineering of Aureobasidium 
            pullulans.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 27:2025.01.25.634885. doi: 
            10.1101/2025.01.25.634885.
  PUBMED    39975359
REFERENCE   2  (bases 1 to 4815)
  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: "bioRxiv 
            [Preprint]. 2025 Jan 27:2025.01.25.634885. doi: 
            10.1101/2025.01.25.634885."
COMMENT     Sequence Label: pAPInt-mScarlet-G418
FEATURES             Location/Qualifiers
     source          1..4815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     847..866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        847..865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(909..1106)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     terminator      complement(2330..2517)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     2438..2462
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        complement(3316..3334)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3317..3334)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3415..3434)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3534..3556
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3594..3612)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3680..3784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3785..3853,3854..4645)
                     /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(4003..4022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggttaa cctggcttat
      841 cgaaattaat acgactcact atagggagac cggcagatcc gcggccgcat aggccactag
      901 tggatctgca gtatagcgac cagcattcac atacgattga cgcatgatat tactttctgc
      961 gcacttaact tcgcatctgg gcagatgatg tcgaggcgaa aaaaaatata aatcacgcta
     1021 acatttgatt aaaatagaac aactacaata taaaaaaact atacaaatga caagttcttg
     1081 aaaacaagaa tctttttatt gtcagtactg actagaagaa ctcgtcgagc atgaggtgga
     1141 actggagctt gttcatgtcc gggttgtcga tgccgtactt ctggaagagg cgcttctgga
     1201 gcgacggcga gaactcgccg aggcagttcc agaggatggc gaggtcctgg tagcggtcgg
     1261 cgatgccgac gcggccgacg tcgatgcagc cgatgagctt gccctcgtcg aagatgaggt
     1321 tgtcgagcga gaagtcgccg tgcgtgacga ccgagtccgg cgagaacggg aggagcttgt
     1381 gcatctcctt ccagacctgc tcgaccggcc agccgttgcg ctcgtcgtcg aagtccgagg
     1441 cgtcgacgag gccgttgttc atgcgcgact gggcctgggc gaggcggaag acgcggtccg
     1501 agttgaacgg gcagttgcag accgggatcg agtggaggcg gcggaggaag acggcgaggg
     1561 cgtcgacgat gttctcgccc gagtccgggt actcctcgag gacctggaag gccgtcttgc
     1621 ccgggatggc cgtcgtgagg agccaggcgt cgtccggcgt gcggatgaag tgcttgatcg
     1681 tcgggagcgg catgaactcc gtgagccagt tgaggcggac catctcgtcc gtgacgtcgt
     1741 tggcgaccga gcccttgccg tgcttgagga agagctccgg ggcgtccggc ttgccgtaga
     1801 ggcggtagat cgtggcgccc gactggccga cgttgtcgcg ggaccacttg tagccgtaga
     1861 ggtcggcgtc catgttcgag ttgaggcgcg ggcgcgagac gtgcgtcttc tccttgccca
     1921 tgaattctgc tgtgtaaggt gtgcgtgaga gagagggtcg tgagtgagtc ttgaagagaa
     1981 acgtcaaagc ttgaagcgta gcagaatacg gaaaacgact tgggggaatg gcgaggagcg
     2041 tcgcctttta tgctccaagc ggagacgatg aggccgatat tacagtggct tagtagcgtg
     2101 tacaatattt ccataaaagg ctacatggtg agtttttcac gtgaccgtaa ttagtcccaa
     2161 ttaggcttta ttgtatattg ctactaagca tcgttaaatg acctgctcga agcttcgtct
     2221 ataaaaggcg ggcgtgtgtg cgcttccccc gctacgtctt tttcacaaac actttgcgac
     2281 ttgaccaaga cctttcaccc gcaccaccct gttatcccta gcggatctgc cggtagaggt
     2341 gtggtcaata agagcgacct catactatac ctgagaaagc aacctgacct acaggaaaga
     2401 gttactcaag aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac
     2461 acttattttt tttataactt atttaataat aaaaatcata aatcataaga aattcgctta
     2521 tttagaagtg gcgctctaga ttcacttgta gagctcgtcc atgccgccgg tagagtggcg
     2581 gccctcagag cgctcgtatt gctcgacgac ggtgtagtcc tcgttgtgag aggtgatgtc
     2641 gagcttgcgg tcgacgttgt aggcgccagg catttggaca ggcttcttgg ccttgtaggt
     2701 ggtcttgaag tcggcgaggt agcggccgcc gtccttgagg cggagggcca tcttgatgtc
     2761 gcccttgagg acgccgtcct cagggtagag gcgctcggta gaggcctccc agcccattgt
     2821 cttcttttgc atgacagggc cgtcaggagg gaagttggtg ccgcggagct tgaccttgta
     2881 gatgagggtg ccgtcctcga gagaggtgtc ttgggtgacg gtgacggcgc cgccgtcctc
     2941 gaagttcatg acgcgctccc acttgaagcc ctcagggaaa gattgcttgt agtagtcagg
     3001 gatgtcggca gggtgcttgg tgaaggcgcg agagccgtac atgaattgag gagagaggat
     3061 gtcccaagag aaagggagag ggccgccctt ggtgaccttg agcttggcgg tttgggtgcc
     3121 ctcgtaaggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcatagagcc
     3181 ctccatgtgg accttgaagc gcatgaactc cttgatgacg gcctcgccct tagagaccat
     3241 ggtaccagaa ccagaaccag caccgtcacc gatccgtcga cctgcagcgt acgaagcttc
     3301 agctggcggc cgcgttctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat
     3361 gtattaattg tagccgcgtt ctaacgacaa tatgtccata tggtgcactc tcagtacaat
     3421 ctgctctgat gccgcatagt taagccagcc ccaacacccg ccaacacccg ctgacgcgcc
     3481 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3541 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3601 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3661 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3721 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3781 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3841 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3901 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3961 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4021 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4081 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4141 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4201 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4261 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4321 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4381 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4441 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4501 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4561 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4621 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4681 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4801 gatcaaagga tcttc
//
