LOCUS       sequence-63302-e        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  AUTHORS   Zordan RE, Ren Y, Pan SJ, Rotondo G, De Las Penas A, Iluore J, 
            Cormack BP
  TITLE     Expression plasmids for use in Candida glabrata.
  JOURNAL   G3 (Bethesda). 2013 Oct 3;3(10):1675-86. doi: 10.1534/g3.113.006908.
  PUBMED    23934995
REFERENCE   2  (bases 1 to 6534)
  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"; doi: "10.1534/g3.113"; 
            journalName: "G3 (Bethesda)"; date: "2013-10-3- 3"; volume: "3"; 
            issue: "10"; pages: "1675-86"
COMMENT     Sequence Label: pGRB2.3
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        666..737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(738..806,807..1598)
                     /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(956..975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2258..2277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2511..2528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2681..2686,2687..2704,2705..2711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2735
                     /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     2724..2746
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2743..2759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2743..2759
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2778..2798
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2780..2798
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3368..3384
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3368..3384
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3404..4120
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4550..4569)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4551..4569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4579..4596)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4579..4595)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4588..4610)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4736..5191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4823..4842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5033..5054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5322..6125)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6126..6341)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6365..6384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6484..6506
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt
      121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag
      181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt
      241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat
      301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt
      361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa
      421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac
      481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt
      541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca
      601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat
      661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2641 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctccata
     2821 aagcacgtgg cctcttatcg agaaagaaat taccgtcgct cgtgatttgt ttgcaaaaag
     2881 aacaaaactg aaaaaaccca gacacgctcg acttcctgtc ttcctattga ttgcagcttc
     2941 caatttcgtc acacaacaag gtcctagcga cggctcacag gttttgtaac aagcaatcga
     3001 aggttctgga atggcgggaa agggtttagt accacatgct atgatgccca ctgtgatctc
     3061 cagagcaaag ttcgttcgat cgtactgtta ctctctccct ttcaaacaga attgtccgaa
     3121 tcgtgtgaca acaacagcct gttctcacac actcttttct tctaaccaag ggggtggttt
     3181 agtttagtag aacctcgtga aacttacatt tacatatata taaacttgca taaattggtc
     3241 aatgcaagaa atacatattt ggtcttttct aattcgtagt ttttcaagtt cttagatgct
     3301 ttctttttct cttttttaca gatcatcaag gaagtaatta tctacttttt acaacaaata
     3361 taaaacatct agaactagtg gatcccccgg gctgcaggaa ttcatgtcta aaggtgaaga
     3421 attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa
     3481 attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt
     3541 tatttgtact actggtaaat tgccggttcc atggccaacc ttagtcacta ctttcggtta
     3601 tggtgttcaa tgttttgcta gatacccaga tcatatgaaa caacatgact ttttcaagtc
     3661 tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta
     3721 caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa
     3781 aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa
     3841 ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa
     3901 aattagacac aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac
     3961 tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc
     4021 cttatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc
     4081 tgctggtatt acccatggta tggatgaatt gtacaaataa ctgcaggtcg acctcgagaa
     4141 cacaacccac agctaccacc atcaacaata tttatatata taacgtacac atagaaatca
     4201 cacaaacaga gtatttattc ttaactacat gaactaccat cagaccgtct gggccactat
     4261 ataatgtgcc attcataaac gtgatcactt tacgtagcag gcaaccccag gtgaaaattt
     4321 ttcagcgagc tgccagattg tcaggtgaaa aactgaaaaa aacttctggg cgatgagctt
     4381 gtggcgggaa attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta
     4441 ttcttgtaat atttggaaga atagaaaatc aagactactg cctttctttt catattatag
     4501 aggaagaaat acgcacgaac acgatataga ggtaaaggta cccaattcgc cctatagtga
     4561 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     4621 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     4681 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     4741 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4801 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4861 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4921 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4981 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5041 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5101 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5161 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     5221 tgtgcggtat ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt
     5281 tgtgagttta gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat
     5341 cttctcaaat atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc
     5401 cttccctttg caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca
     5461 tcatccacgg ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg
     5521 ggtgtcataa tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata
     5581 aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct
     5641 ccagtagata gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc
     5701 tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg
     5761 tgtgcattcg taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat
     5821 ttgactgtat taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg
     5881 gcggataatg cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc
     5941 acatgtgttt ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc
     6001 ttggtggtac gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta
     6061 aatagcttgg cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc
     6121 gacatgattt atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact
     6181 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     6241 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     6301 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc
     6361 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6421 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6481 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
