LOCUS       sequence-468366-        4820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for deleting or C-terminally tagging endogenous genes with 
            GFP and selection on G418..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4820)
  AUTHORS   Colarusso AV, Williams AM, Gladfelter AS, Wirshing ACE, Lew DJ
  TITLE     Optimized vectors for genetic engineering of Aureobasidium 
            pullulans.
  JOURNAL   Mol Biol Cell. 2025 Jun 1;36(6):mr5. doi: 10.1091/mbc.E25-02-0059. 
            Epub 2025 Apr 9.
  PUBMED    40202829
REFERENCE   2  (bases 1 to 4820)
  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: "Mol Biol 
            Cell."; date: "2025-06-1"; volume: "36(6):mr5. doi"; pages: " 
            10.1091/mbc.E25-02-0059. Epub 2025 Apr 9"
COMMENT     Sequence Label: pAPInt-GFP-G418
FEATURES             Location/Qualifiers
     source          1..4820
                     /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"
     primer_bind     2929..2948
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(3321..3339)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3322..3339)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3420..3439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3539..3561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3599..3617)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3685..3789
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3790..3858,3859..4650)
                     /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(4008..4027)
                     /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 gcgctcactt gtagagctcg tccatgccgt gggtgatgcc ggcggcggtg
     2581 acgaactcga ggaggaccat gtggtcgcgc ttctcgttag ggtccttaga gagggcagat
     2641 tgggtagaga ggtagtggtt gtcagggagg aggacagggc cgtcgccgat aggggtgttt
     2701 tgttggtagt ggtcggcgag ttggacagag ccgtcctcga tgttgtggcg gatcttgaag
     2761 ttgaccttga tgccgttctt ttgcttgtcg gccatgatgt agacgttgtg agagttgtag
     2821 ttgtactcga gcttgtggcc gaggatgttg ccgtcctcct tgaagtcgat gcccttgagc
     2881 tcgatgcggt tgacgagggt gtcgccctcg aacttgacct cggcgcgggt cttgtagttg
     2941 ccgtcgtcct tgaagaagat ggtgcgctct tggacgtagc cctcaggcat ggcagacttg
     3001 aagaagtcgt gttgcttcat gtggtcaggg tagcgggcga agcattggac gccgtagccg
     3061 aaggtggtga cgagggtagg ccaagggaca gggagcttgc cggtggtgca gatgaacttg
     3121 agggtgagct tgccgtaggt ggcgtcgccc tcgccctcgc cagagacaga gaacttgtgg
     3181 ccgttgacgt cgccgtcgag ctcgacgagg atagggacga cgccggtgaa gagctcctcg
     3241 cccttagaca tagaaccaga accagcaccg tcaccgatcc gtcgacctgc agcgtacgaa
     3301 gcttcagctg gcggccgcgt tctatagtgt cacctaaatc gtatgtgtat gatacataag
     3361 gttatgtatt aattgtagcc gcgttctaac gacaatatgt ccatatggtg cactctcagt
     3421 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3481 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3541 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3601 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3661 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3721 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3781 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3841 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3901 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3961 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4021 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4081 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4141 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4201 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4261 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4321 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4381 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4441 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4501 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4561 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4621 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4681 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4741 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4801 gaaaagatca aaggatcttc
//
