LOCUS       sequence-370819-        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid without an ARS sequence for in vivo gene amplification 
            (HamAmp) at RPL25 locus (~12 copy).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 7686)
  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: pILGFP3AG4ARSd
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2455..2472)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2455..2471)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2464..2486)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2680..2699)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2799..2821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2859..2877)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2945..3049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3050..3118,3119..3910)
                     /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(3268..3287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4081..4669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4570..4589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4823..4840
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4957..4978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4993..4998,4999..5016,5017..5023)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5031..5047
                     /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)."
     primer_bind     5036..5058
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5055..5071
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5055..5071
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(6351..7061)
                     /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"
     promoter        complement(7084..7497)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ctcacgaaga agttaagctt gagcatcgga ccgaagcata agaatgcagc aaattgataa
       61 tattcccgtt aagccagtcg taggaaaagg aaaagaatga gtagcaaatg tcacacaaca
      121 aacgcgcacc atccctacat acacaaacac acacacacac acacatatat atatattcta
      181 ccccatataa gcgagtgatt catcatgaaa agaatttatc acttcgaatt taaataataa
      241 cattcttacc tgatgtacaa ctctttactc tcatcttgcg aaaagaaaag ggcagggtgc
      301 cgctgaaaca gcggcgttgc tgatgaaaat aaccctgtgt tttcgcctca ttctctcaaa
      361 accttcacag atccttttct tgttatagcc tcaataaaat cacagtacac taacacgtaa
      421 ttataaatag taaccggaac cctcgctgca atacgcaaat tggtacctac tttgtaaggc
      481 acaggaaacc ccgtgcactc tgccactaca ctttatttac gcttaatcat gaatttaaca
      541 tgttatgaaa gatttattgt atatttaaag aaaaatttaa aaataatatt aaatttatta
      601 attaaaccaa ttagagctag ctttgggccc gcaggctaac cggaacctgt attatttagt
      661 ttatgctacg ttaaataaag acctttcgtt cacataactg aatgtgtaat ggccttgaga
      721 tttcaagcat accaagttgg tggagacggg gtcgttacaa aagactcttt aagccaagat
      781 ttccttgaca gccttggcga tagcatcgcc aacctcagtg gtagagttag aaccaccaag
      841 gtcaccggtt ctgacacctg catccaagac atttctaaca gcttcttcaa gagccctacc
      901 ttcttcaacc aaatccaagg ataacttcaa catcatagct gcagataaga tggtagcaat
      961 tgggttaacc ttgtttgctg gtaaatctgg ggcagaacca tgacatggtt cgtacaaacc
     1021 gaatgccttg ttagtgtcag gtagggaagc tagagatgca gaaggtaata aacccaaaga
     1081 acctggaata acagaggctt catcggagat aatatcacca aacatgttgt tggtaataac
     1141 aacaccgttt agcttagttg gtgatttaac caaaatcata gcagcagagt cgatcaattg
     1201 gtgctgaaca gttaattgtg ggaactcagt cttgatggtt tcttcaacag tctttctcca
     1261 caatctggaa gaggcaagca cgttagcctt gtcaagagac cagattggta atggtgggtt
     1321 ttgttgcaat gccaagaaag cagccattct tgtaattctt tgaacttcag gaacactgta
     1381 tttctcagag tcccaagcaa ctccgtcacc ttcatcttct tttctttcac caaagtagat
     1441 accaccaacc aattctctaa cgacgacgaa atcggtaccc tttgcatatt caggcttcaa
     1501 aggagaaaga tctagtaaag aatcagaagc aaagttacat ggtctcaagt tggcgtatag
     1561 acccaattcc tttctgatct tcaatagacc ttgttctggt ctaactgcgc ccgtacccca
     1621 ttttggacca ccaacagcac ctagtaagac agcatcggct ttcttagagg cttctagagc
     1681 ttcatctggt aaaggagtgc cagtggcatc gatggcagca cccccgatca agtgatgttg
     1741 gaaattgaac ttaatttctg gacggacttc agcaatggca ttcaagacct taatagcttc
     1801 gtcagtaact tctttaccga cgtgatcacc cggtaggaca acgatattct tagacatttt
     1861 gcggccgctt tatcttattg atcttctttg tttagccttt tcaacagcag aagaaaagca
     1921 tatgagatac atttaaaatt gttttaaaat aattttaaaa gataatatct aaaagattgt
     1981 atgcatacga acgaagttgc attgggaagc cccaacgcag tgaggcaagc cgaaggtaga
     2041 aagaaaatcc ccttgtgcgt ccagtgagaa acagcaagca gtgggtagga aggcccagtt
     2101 tagcctgact gttccaacgg ggctcccagg ctgaggaaat gcttctctcc gtaatgggag
     2161 agcctaactc cagagcgtgg acttcggctc acggttttct ctttttttgg cgcacaatga
     2221 ctgctgagac aaaaaaagaa agcaactcca tgtaaacacc cgtacatatt aaataagtga
     2281 cccatgcctc ctcatctctt cattgcaaag tacggaaacc gttagattgg taccaacaag
     2341 gttgctatat cgggcataca agataagaaa cttaaagaac gccgagacaa gaatatgtaa
     2401 tgataagcct cctccggggt gttagactga ttagtacatg tttaaactga attcactggc
     2461 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     2521 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2581 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     2641 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     2701 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2761 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2821 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     2881 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2941 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3001 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3061 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3121 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3181 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3241 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3301 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3361 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3421 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3481 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3541 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3601 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3661 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3721 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3781 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3841 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3901 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4081 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4141 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4201 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4261 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4321 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4381 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4441 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4501 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4561 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4621 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4681 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4741 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4801 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4861 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4921 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     4981 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5041 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg tttaaactat
     5101 gttccttgat acctcgacgt ggagcacgaa atagaagaaa aaagtataat gaggatggaa
     5161 taattttctt gaatctgagc agaaaaacga acgaacactc taatggattt gttaatggct
     5221 gttggtagta aaaagaacaa cttccacagc atcaacgctt ctatcaaaac gattgctaat
     5281 aaactgttga gcccgataat tacgttgctc accatatctt aatcaagtcc ttaatgcact
     5341 attcttgtcg tcggatagtt tatttacgtg tggttttcgt atctcatcaa tatcaactct
     5401 actagttccc atctatcttc ctctcaattt tctcgtgcta ttaatattgc agttcacata
     5461 ccagatggag ccattggcac aaatgtggtt tcctttctta ttgctgtatc caacttttgt
     5521 gaagctacag gtgcgacaaa cataaaaaag ttcttgccgt taagaagctg aagttgcttc
     5581 tcgctatacc agaggggatt attgtcggga gtcatgatgc gacatttacc cggttgagta
     5641 aggaaggggt ggaggaatca gaagagagat aaacaaacat acacgtcaaa acagtatagt
     5701 tcaagtaaca catatagaca tgtacacgcg cgattacgtg tgtacgtgca catgttaatg
     5761 ttatatacat atatagatat agatatataa ttgcaataat gaactacaaa ttattatacc
     5821 cccgttcttt tttttttctt ttttttagta atttttcttt agatttatcg gaatacttac
     5881 tttggtgctt caccttcggc ggcagcaggt ggctgctgtt gctgatgttg ttgttgttgt
     5941 tgttggtctt cagcttgacc ggactcggac atgtctgaag tccataaggt taaattgtcc
     6001 cttagcagtt gcatgataag tgtgctatct ttgtatgatt cttcagacag agtgtccaac
     6061 tcagcaatag cgtcgtcaaa agcttgcttg gccaaatggc aggctttgtc tggagagttt
     6121 tgaatttcgc atgcaaatca ttacgaccga gattcccggg taataactga tataattaaa
     6181 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     6241 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     6301 ctaccttagc atcccttccc tttgcaaata gtcctactag tttaagatct tttgtacaat
     6361 tcatccatac catgggtaat accagcagca gtaacaaatt ctaacaagac catgtggtct
     6421 ctcttttcgt ttggatcttt ggataaggca gattgagtgg ataagtaatg gttgtctggt
     6481 aacaagactg gaccatcacc aattggagta ttttgttgat aatggtcagc taattgaaca
     6541 gaaccatctt caatgttgtg tctaattttg aagttaactt tgataccatt cttttgtttg
     6601 tcagccatga tgtaaacatt gtgagagtta tagttgtatt ccaatttgtg acctaaaatg
     6661 ttaccatctt ctttaaaatc aatacctttt aattcgattc tattaactaa ggtatcacct
     6721 tcaaacttga cttcagctct ggtcttgtag ttaccgtcat ctttgaaaaa aatagttctt
     6781 tcttgaacat aaccttctgg catggcagac ttgaaaaagt catgttgttt catatgatct
     6841 gggtatctcg caaaacattg aacaccataa ccgaaagtag tgactaaggt tggccatgga
     6901 actggcaatt taccagtagt acaaataaat tttaaggtca atttaccgta agtagcatca
     6961 ccttcacctt caccggagac agaaaatttg tgaccattaa catcaccatc taattcaacc
     7021 aaaattggga caacaccagt gaataattct tcacctttag aggatcccat tttgtaatta
     7081 aaacttagat tagattgcta tgctttcttt ctaatgagca agaagtaaaa aaagttgtaa
     7141 tagaacaaga aaaatgaaac tgaaacttga gaaattgaag accgtttatt aacttaaata
     7201 tcaatgggag gtcatcgaaa gagaaaaaaa tcaaaaaaaa aaattttcaa gaaaaagaaa
     7261 cgtgataaaa atttttattg cctttttcga cgaagaaaaa gaaacgaggc ggtctctttt
     7321 ttcttttcca aacctttagt acgggtaatt aacgacaccc tagaggaaga aagaggggaa
     7381 atttagtatg ctgtgcttgg gtgttttgaa gtggtacggc gatgcgcgga gtccgagaaa
     7441 atctggaaga gtaaaaaagg agtagaaaca ttttgaagct atggtgtgtg ggggatcact
     7501 tgtgggggat tgggtgtgat gtaaggattc gcggtcctcg aaaattaaaa gtccaacgcg
     7561 cctgttgctt cctatgtgat atgtattata tgtaatatgc ataaatatat ctactgcatt
     7621 gtattttgaa cgtacaaagt atgcattgtt tatacgcctg caggttaatt tatctcatga
     7681 agtaac
//
