LOCUS       sequence-370815-        6923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A parental plasmid for in vivo gene amplification at RPL25 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6923)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 6923)
  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"
COMMENT     Sequence Label: pILGFP1DFB
FEATURES             Location/Qualifiers
     source          1..6923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..20
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4..20
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2692..2725)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2936..3349
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             3372..4082
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(4327..4343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4327..4343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4340..4362)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4351..4367
                     /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)."
     promoter        complement(join(4375..4381,4382..4399,4400..4405))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4420..4441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4558..4575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4729..5317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4809..4828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5488..6279,6280..6348))
                     /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     6111..6130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6349..6453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6521..6539
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6577..6599)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6699..6718
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6912..6923,1..11)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gttgtaaaac gacggccagt gaattcagtt taaacatgta ctaatcagtc taacaccccg
       61 gaggaggctt atcattacat attcttgtct cggcgttctt taagtttctt atcttgtatg
      121 cccgatatag caaccttgtt ggtaccaatc taacggtttc cgtactttgc aatgaagaga
      181 tgaggaggca tgggtcactt atttaatatg tacgggtgtt tacatggagt tgctttcttt
      241 ttttgtctca gcagtcattg tgcgccaaaa aaagagaaaa ccgtgagccg aagtccacgc
      301 tctggagtta ggctctccca ttacggagag aagcatttcc tcagcctggg agccccgttg
      361 gaacagtcag gctaaactgg gccttcctac ccactgcttg ctgtttctca ctggacgcac
      421 aaggggattt tctttctacc ttcggcttgc ctcactgcgt tggggcttcc caatgcaact
      481 tcgttcgtat gcatacaatc ttttagatat tatcttttaa aattatttta aaacaatttt
      541 aaatgtatct catatgcttt tcttctgctg ttgaaaaggc taaacaaaga agatcaataa
      601 gataaagcgg ccgcaaaatg tctaagaata tcgttgtcct accgggtgat cacgtcggta
      661 aagaagttac tgacgaagct attaaggtct tgaatgccat tgctgaagtc cgtccagaaa
      721 ttaagttcaa tttccaacat cacttgatcg ggggtgctgc catcgatgcc actggcactc
      781 ctttaccaga tgaagctcta gaagcctcta agaaagccga tgctgtctta ctaggtgctg
      841 ttggtggtcc aaaatggggt acgggcgcag ttagaccaga acaaggtcta ttgaagatca
      901 gaaaggaatt gggtctatac gccaacttga gaccatgtaa ctttgcttct gattctttac
      961 tagatctttc tcctttgaag cctgaatatg caaagggtac cgatttcgtc gtcgttagag
     1021 aattggttgg tggtatctac tttggtgaaa gaaaagaaga tgaaggtgac ggagttgctt
     1081 gggactctga gaaatacagt gttcctgaag ttcaaagaat tacaagaatg gctgctttct
     1141 tggcattgca acaaaaccca ccattaccaa tctggtctct tgacaaggct aacgtgcttg
     1201 cctcttccag attgtggaga aagactgttg aagaaaccat caagactgag ttcccacaat
     1261 taactgttca gcaccaattg atcgactctg ctgctatgat tttggttaaa tcaccaacta
     1321 agctaaacgg tgttgttatt accaacaaca tgtttggtga tattatctcc gatgaagcct
     1381 ctgttattcc aggttctttg ggtttattac cttctgcatc tctagcttcc ctacctgaca
     1441 ctaacaaggc attcggtttg tacgaaccat gtcatggttc tgccccagat ttaccagcaa
     1501 acaaggttaa cccaattgct accatcttat ctgcagctat gatgttgaag ttatccttgg
     1561 atttggttga agaaggtagg gctcttgaag aagctgttag aaatgtcttg gatgcaggtg
     1621 tcagaaccgg tgaccttggt ggttctaact ctaccactga ggttggcgat gctatcgcca
     1681 aggctgtcaa ggaaatcttg gcttaaagag tcttttgtaa cgaccccgtc tccaccaact
     1741 tggtatgctt gaaatctcaa ggccattaca cattcagtta tgtgaacgaa aggtctttat
     1801 ttaacgtagc ataaactaaa taatacaggt tccggttagc ctgcgggccc aaagctagct
     1861 ctaattggtt taattaataa atttaatatt atttttaaat ttttctttaa atatacaata
     1921 aatctttcat aacatgttaa attcatgatt aagcgtaaat aaagtgtagt ggcagagtgc
     1981 acggggtttc ctgtgcctta caaagtaggt accaatttgc gtattgcagc gagggttccg
     2041 gttactattt ataattacgt gttagtgtac tgtgatttta ttgaggctat aacaagaaaa
     2101 ggatctgtga aggttttgag agaatgaggc gaaaacacag ggttattttc atcagcaacg
     2161 ccgctgtttc agcggcaccc tgcccttttc ttttcgcaag atgagagtaa agagttgtac
     2221 atcaggtaag aatgttatta tttaaattcg aagtgataaa ttcttttcat gatgaatcac
     2281 tcgcttatat ggggtagaat atatatatat gtgtgtgtgt gtgtgtgttt gtgtatgtag
     2341 ggatggtgcg cgtttgttgt gtgacatttg ctactcattc ttttcctttt cctacgactg
     2401 gcttaacggg aatattatca atttgctgca ttcttatgct tcggtccgat gctcaagctt
     2461 aacttcttcg tgaggaagga aagtgttgtc tcctactttt ttcaaatttt cgaattgtat
     2521 ttatatttat ttagtacttc ttgagtttac atatccttcg taaaaatgca acttttgtcg
     2581 aaaaacactt ccaaaaaaaa ataataatga atttatgaag catactaacg agcgagcaca
     2641 tcgctgaggt atcattactt catgagataa attaactcga gctcgagcct aataacttcg
     2701 tatagcatac attatacgaa gttatattaa gggttgctcg accagaaagc gaccacccaa
     2761 ctttggctga taatagcgta taaacaatgc atactttgta cgttcaaaat acaatgcagt
     2821 agatatattt atgcatatta catataatac atatcacata ggaagcaaca ggcgcgttgg
     2881 acttttaatt ttcgaggacc gcgaatcctt acatcacacc caatccccca caagtgatcc
     2941 cccacacacc atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg
     3001 actccgcgca tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc
     3061 tttcttcctc tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga
     3121 ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt
     3181 tcttgaaaat tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag
     3241 ttaataaacg gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact
     3301 ttttttactt cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca
     3361 aaatgggatc ctctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat
     3421 tagatggtga tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta
     3481 cttacggtaa attgacctta aaatttattt gtactactgg taaattgcca gttccatggc
     3541 caaccttagt cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata
     3601 tgaaacaaca tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta
     3661 tttttttcaa agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata
     3721 ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag
     3781 gtcacaaatt ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa
     3841 agaatggtat caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat
     3901 tagctgacca ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca
     3961 accattactt atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca
     4021 tggtcttgtt agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca
     4081 aaagatctta aactagtagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     4141 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     4201 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     4261 tatcagttat tacccgggaa tctcggtcgt aatgatttaa atgcatgcaa gcttggcgta
     4321 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4381 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4441 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4501 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4561 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4621 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4681 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4741 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4801 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4861 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4921 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4981 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5041 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5101 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5161 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5221 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5281 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5341 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5401 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5461 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5521 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5581 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5641 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5701 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5761 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5821 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5881 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5941 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6001 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6061 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6121 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6181 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6241 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6301 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6361 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6421 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6481 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     6541 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     6601 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     6661 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     6721 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6781 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6841 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6901 gccagggttt tcccagtcac gac
//
