LOCUS       sequence-370816-        7889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid for in vivo gene amplification (HamAmp) at RPL25 locus (1 
            copy reference).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7889)
  TITLE     Vickers Lab plasmids
REFERENCE   2  (bases 1 to 7889)
  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: pILGFP3A5C
FEATURES             Location/Qualifiers
     source          1..7889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..50)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(47..69)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    58..74
                     /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(82..88,89..106,107..112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(265..282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(436..1024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(516..535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1195..1986,1987..2055))
                     /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     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2228..2246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2284..2306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2406..2425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2619..2641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2633..2650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2634..2650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5322..5355)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5566..5979
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             6002..6712
                     /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 gaacatagtt taaacaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
       61 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2221 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2461 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2641 gacggccagt gaattcagtt taaacatgta ctaatcagtc taacaccccg gaggaggctt
     2701 atcattacat attcttgtct cggcgttctt taagtttctt atcttgtatg cccgatatag
     2761 caaccttgtt ggtaccaatc taacggtttc cgtactttgc aatgaagaga tgaggaggca
     2821 tgggtcactt atttaatatg tacgggtgtt tacatggagt tgctttcttt ttttgtctca
     2881 gcagtcattg tgcgccaaaa aaagagaaaa ccgtgagccg aagtccacgc tctggagtta
     2941 ggctctccca ttacggagag aagcatttcc tcagcctggg agccccgttg gaacagtcag
     3001 gctaaactgg gccttcctac ccactgcttg ctgtttctca ctggacgcac aaggggattt
     3061 tctttctacc ttcggcttgc ctcactgcgt tggggcttcc caatgcaact tcgttcgtat
     3121 gcatacaatc ttttagatat tatcttttaa aattatttta aaacaatttt aaatgtatct
     3181 catatgcttt tcttctgctg ttgaaaaggc taaacaaaga agatcaataa gataaagcgg
     3241 ccgcaaaatg tctaagaata tcgttgtcct accgggtgat cacgtcggta aagaagttac
     3301 tgacgaagct attaaggtct tgaatgccat tgctgaagtc cgtccagaaa ttaagttcaa
     3361 tttccaacat cacttgatcg ggggtgctgc catcgatgcc actggcactc ctttaccaga
     3421 tgaagctcta gaagcctcta agaaagccga tgctgtctta ctaggtgctg ttggtggtcc
     3481 aaaatggggt acgggcgcag ttagaccaga acaaggtcta ttgaagatca gaaaggaatt
     3541 gggtctatac gccaacttga gaccatgtaa ctttgcttct gattctttac tagatctttc
     3601 tcctttgaag cctgaatatg caaagggtac cgatttcgtc gtcgttagag aattggttgg
     3661 tggtatctac tttggtgaaa gaaaagaaga tgaaggtgac ggagttgctt gggactctga
     3721 gaaatacagt gttcctgaag ttcaaagaat tacaagaatg gctgctttct tggcattgca
     3781 acaaaaccca ccattaccaa tctggtctct tgacaaggct aacgtgcttg cctcttccag
     3841 attgtggaga aagactgttg aagaaaccat caagactgag ttcccacaat taactgttca
     3901 gcaccaattg atcgactctg ctgctatgat tttggttaaa tcaccaacta agctaaacgg
     3961 tgttgttatt accaacaaca tgtttggtga tattatctcc gatgaagcct ctgttattcc
     4021 aggttctttg ggtttattac cttctgcatc tctagcttcc ctacctgaca ctaacaaggc
     4081 attcggtttg tacgaaccat gtcatggttc tgccccagat ttaccagcaa acaaggttaa
     4141 cccaattgct accatcttat ctgcagctat gatgttgaag ttatccttgg atttggttga
     4201 agaaggtagg gctcttgaag aagctgttag aaatgtcttg gatgcaggtg tcagaaccgg
     4261 tgaccttggt ggttctaact ctaccactga ggttggcgat gctatcgcca aggctgtcaa
     4321 ggaaatcttg gcttaaagag tcttttgtaa cgaccccgtc tccaccaact tggtatgctt
     4381 gaaatctcaa ggccattaca cattcagtta tgtgaacgaa aggtctttat ttaacgtagc
     4441 ataaactaaa taatacaggt tccggttagc ctgcgggccc aaagctagct ctaattggtt
     4501 taattaataa atttaatatt atttttaaat ttttctttaa atatacaata aatctttcat
     4561 aacatgttaa attcatgatt aagcgtaaat aaagtgtagt ggcagagtgc acggggtttc
     4621 ctgtgcctta caaagtaggt accaatttgc gtattgcagc gagggttccg gttactattt
     4681 ataattacgt gttagtgtac tgtgatttta ttgaggctat aacaagaaaa ggatctgtga
     4741 aggttttgag agaatgaggc gaaaacacag ggttattttc atcagcaacg ccgctgtttc
     4801 agcggcaccc tgcccttttc ttttcgcaag atgagagtaa agagttgtac atcaggtaag
     4861 aatgttatta tttaaattcg aagtgataaa ttcttttcat gatgaatcac tcgcttatat
     4921 ggggtagaat atatatatat gtgtgtgtgt gtgtgtgttt gtgtatgtag ggatggtgcg
     4981 cgtttgttgt gtgacatttg ctactcattc ttttcctttt cctacgactg gcttaacggg
     5041 aatattatca atttgctgca ttcttatgct tcggtccgat gctcaagctt aacttcttcg
     5101 tgaggaagga aagtgttgtc tcctactttt ttcaaatttt cgaattgtat ttatatttat
     5161 ttagtacttc ttgagtttac atatccttcg taaaaatgca acttttgtcg aaaaacactt
     5221 ccaaaaaaaa ataataatga atttatgaag catactaacg agcgagcaca tcgctgaggt
     5281 atcattactt catgagataa attaactcga gctcgagcct aataacttcg tatagcatac
     5341 attatacgaa gttatattaa gggttgctcg accagaaagc gaccacccaa ctttggctga
     5401 taatagcgta taaacaatgc atactttgta cgttcaaaat acaatgcagt agatatattt
     5461 atgcatatta catataatac atatcacata ggaagcaaca ggcgcgttgg acttttaatt
     5521 ttcgaggacc gcgaatcctt acatcacacc caatccccca caagtgatcc cccacacacc
     5581 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     5641 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     5701 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     5761 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     5821 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
     5881 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
     5941 cttgctcatt agaaagaaag catagcaatc taatctaagt tttaattaca aaatgggatc
     6001 ctctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga
     6061 tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa
     6121 attgacctta aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt
     6181 cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata tgaaacaaca
     6241 tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa
     6301 agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa
     6361 tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt
     6421 ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat
     6481 caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca
     6541 ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt
     6601 atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt
     6661 agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca aaagatctta
     6721 aactagtagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     6781 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     6841 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     6901 tacccgggaa tctcggtcgt aatgatttaa atgcatacat aacattttaa gataagcaag
     6961 tgaatgttga ttcccgtttc ttagtcaaca cttctttcta ttttacccgg tcgttaccct
     7021 attaaaaaaa caacttacaa tcattgttcg ccccttccat acttactgcc actcgcaaaa
     7081 gggcccaacc agggcaatta cgtatcaaaa aatcatgaca ggctgggtaa taaatattcg
     7141 tgaagaaaga agaaattaaa aaaagaaacg aagaagcaaa aaaaagaaaa gactccgttt
     7201 aatcactttc aaccgcggtt tatccggccc cacccatgca taaccctaaa ttattagatc
     7261 acttagcacg tgaaaaagaa acgtttttaa tgtttttttt tttttttctt tttctttttt
     7321 tgcgttggtg aaaatttttt cgcttcctcg agtataatta tctcatctca tctttcatat
     7381 aagataagaa gttttataaa aaccttttgc atcaaaattt tgtagaatat ctctttttct
     7441 tacgctctct ttctttcctt aattgttttc taaagaaccg tgtatttttc tagttcgaat
     7501 ccatcgataa cattaaaaga tggctccatc tggtatgtga actgcaatat taatagcacg
     7561 agaaaattga gaggaagata gatgggaact agtagagttg atattgatga gatacgaaaa
     7621 ccacacgtaa ataaactatc cgacgacaag aatagtgcat taaggacttg attaagatat
     7681 ggtgagcaac gtaattatcg ggctcaacag tttattagca atcgttttga tagaagcgtt
     7741 gatgctgtgg aagttgttct ttttactacc aacagccatt aacaaatcca ttagagtgtt
     7801 cgttcgtttt tctgctcaga ttcaagaaaa ttattccatc ctcattatac ttttttcttc
     7861 tatttcgtgc tccacgtcga ggtatcaag
//
