LOCUS       sequence-369502-        7626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid for in vivo gene amplification (HamAmp) at RPL25 locus 
            (~47 copy).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 7626)
  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: pILGFP3AA5
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(20..36)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(20..36)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(33..55)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    44..60
                     /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(68..74,75..92,93..98))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(251..268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(422..1010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(502..521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1181..1972,1973..2041))
                     /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     1804..1823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2042..2146)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2214..2232
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2270..2292)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2392..2411
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2605..2627
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2619..2636
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2620..2636
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5308..5341)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5552..5965
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             5988..6698
                     /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"
ORIGIN
        1 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
       61 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
      121 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
      181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      301 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      361 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      601 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      841 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1081 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2161 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2221 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2281 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2341 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2401 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2461 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2521 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2581 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2641 tcagtttaaa catgtactaa tcagtctaac accccggagg aggcttatca ttacatattc
     2701 ttgtctcggc gttctttaag tttcttatct tgtatgcccg atatagcaac cttgttggta
     2761 ccaatctaac ggtttccgta ctttgcaatg aagagatgag gaggcatggg tcacttattt
     2821 aatatgtacg ggtgtttaca tggagttgct ttcttttttt gtctcagcag tcattgtgcg
     2881 ccaaaaaaag agaaaaccgt gagccgaagt ccacgctctg gagttaggct ctcccattac
     2941 ggagagaagc atttcctcag cctgggagcc ccgttggaac agtcaggcta aactgggcct
     3001 tcctacccac tgcttgctgt ttctcactgg acgcacaagg ggattttctt tctaccttcg
     3061 gcttgcctca ctgcgttggg gcttcccaat gcaacttcgt tcgtatgcat acaatctttt
     3121 agatattatc ttttaaaatt attttaaaac aattttaaat gtatctcata tgcttttctt
     3181 ctgctgttga aaaggctaaa caaagaagat caataagata aagcggccgc aaaatgtcta
     3241 agaatatcgt tgtcctaccg ggtgatcacg tcggtaaaga agttactgac gaagctatta
     3301 aggtcttgaa tgccattgct gaagtccgtc cagaaattaa gttcaatttc caacatcact
     3361 tgatcggggg tgctgccatc gatgccactg gcactccttt accagatgaa gctctagaag
     3421 cctctaagaa agccgatgct gtcttactag gtgctgttgg tggtccaaaa tggggtacgg
     3481 gcgcagttag accagaacaa ggtctattga agatcagaaa ggaattgggt ctatacgcca
     3541 acttgagacc atgtaacttt gcttctgatt ctttactaga tctttctcct ttgaagcctg
     3601 aatatgcaaa gggtaccgat ttcgtcgtcg ttagagaatt ggttggtggt atctactttg
     3661 gtgaaagaaa agaagatgaa ggtgacggag ttgcttggga ctctgagaaa tacagtgttc
     3721 ctgaagttca aagaattaca agaatggctg ctttcttggc attgcaacaa aacccaccat
     3781 taccaatctg gtctcttgac aaggctaacg tgcttgcctc ttccagattg tggagaaaga
     3841 ctgttgaaga aaccatcaag actgagttcc cacaattaac tgttcagcac caattgatcg
     3901 actctgctgc tatgattttg gttaaatcac caactaagct aaacggtgtt gttattacca
     3961 acaacatgtt tggtgatatt atctccgatg aagcctctgt tattccaggt tctttgggtt
     4021 tattaccttc tgcatctcta gcttccctac ctgacactaa caaggcattc ggtttgtacg
     4081 aaccatgtca tggttctgcc ccagatttac cagcaaacaa ggttaaccca attgctacca
     4141 tcttatctgc agctatgatg ttgaagttat ccttggattt ggttgaagaa ggtagggctc
     4201 ttgaagaagc tgttagaaat gtcttggatg caggtgtcag aaccggtgac cttggtggtt
     4261 ctaactctac cactgaggtt ggcgatgcta tcgccaaggc tgtcaaggaa atcttggctt
     4321 aaagagtctt ttgtaacgac cccgtctcca ccaacttggt atgcttgaaa tctcaaggcc
     4381 attacacatt cagttatgtg aacgaaaggt ctttatttaa cgtagcataa actaaataat
     4441 acaggttccg gttagcctgc gggcccaaag ctagctctaa ttggtttaat taataaattt
     4501 aatattattt ttaaattttt ctttaaatat acaataaatc tttcataaca tgttaaattc
     4561 atgattaagc gtaaataaag tgtagtggca gagtgcacgg ggtttcctgt gccttacaaa
     4621 gtaggtacca atttgcgtat tgcagcgagg gttccggtta ctatttataa ttacgtgtta
     4681 gtgtactgtg attttattga ggctataaca agaaaaggat ctgtgaaggt tttgagagaa
     4741 tgaggcgaaa acacagggtt attttcatca gcaacgccgc tgtttcagcg gcaccctgcc
     4801 cttttctttt cgcaagatga gagtaaagag ttgtacatca ggtaagaatg ttattattta
     4861 aattcgaagt gataaattct tttcatgatg aatcactcgc ttatatgggg tagaatatat
     4921 atatatgtgt gtgtgtgtgt gtgtttgtgt atgtagggat ggtgcgcgtt tgttgtgtga
     4981 catttgctac tcattctttt ccttttccta cgactggctt aacgggaata ttatcaattt
     5041 gctgcattct tatgcttcgg tccgatgctc aagcttaact tcttcgtgag gaaggaaagt
     5101 gttgtctcct acttttttca aattttcgaa ttgtatttat atttatttag tacttcttga
     5161 gtttacatat ccttcgtaaa aatgcaactt ttgtcgaaaa acacttccaa aaaaaaataa
     5221 taatgaattt atgaagcata ctaacgagcg agcacatcgc tgaggtatca ttacttcatg
     5281 agataaatta actcgagctc gagcctaata acttcgtata gcatacatta tacgaagtta
     5341 tattaagggt tgctcgacca gaaagcgacc acccaacttt ggctgataat agcgtataaa
     5401 caatgcatac tttgtacgtt caaaatacaa tgcagtagat atatttatgc atattacata
     5461 taatacatat cacataggaa gcaacaggcg cgttggactt ttaattttcg aggaccgcga
     5521 atccttacat cacacccaat cccccacaag tgatccccca cacaccatag cttcaaaatg
     5581 tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt
     5641 caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa
     5701 ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc
     5761 gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt
     5821 tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc
     5881 aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg ctcattagaa
     5941 agaaagcata gcaatctaat ctaagtttta attacaaaat gggatcctct aaaggtgaag
     6001 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
     6061 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
     6121 ttatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttcggtt
     6181 atggtgttca atgttttgcg agatacccag atcatatgaa acaacatgac tttttcaagt
     6241 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
     6301 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
     6361 aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata
     6421 actctcacaa tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca
     6481 aaattagaca caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata
     6541 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg
     6601 ccttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     6661 ctgctggtat tacccatggt atggatgaat tgtacaaaag atcttaaact agtaggacta
     6721 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     6781 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     6841 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgggaatctc
     6901 ggtcgtaatg atttgcatgc ccgccatctc tactcactcc ataatattac atatagatat
     6961 aggacaagcc cgcattttca tactgaaagg taaacttcta ttattatagt ggtatccaac
     7021 gttcaccgct tccagcatag cagaaattac gtgtttttgc atatgttatg ctgatcattg
     7081 tatgcttact accatttttc tttgcttcgc cttgccttct ttgacgtttt tttgaagcaa
     7141 aaaaaaagtc aagacagatg tgcttacaaa accatgtaag gctcattttc aaagaagcta
     7201 ctaatagaaa gagaacaaag cgtttacgag tctggaaaat caatggctcc atctggtatg
     7261 tgaactgcaa tattaatagc acgagaaaat tgagaggaag atagatggga actagtagag
     7321 ttgatattga tgagatacga aaaccacacg taaataaact atccgacgac aagaatagtg
     7381 cattaaggac ttgattaaga tatggtgagc aacgtaatta tcgggctcaa cagtttatta
     7441 gcaatcgttt tgatagaagc gttgatgctg tggaagttgt tctttttact accaacagcc
     7501 attaacaaat ccattagagt gttcgttcgt ttttctgctc agattcaaga aaattattcc
     7561 atcctcatta tacttttttc ttctatttcg tgctccacgt cgaggtatca aggaacatag
     7621 tttaaa
//
