LOCUS       sequence-63299-e        5299 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 5299)
  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 5299)
  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: pGRB2.1
FEATURES             Location/Qualifiers
     source          1..5299
                     /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     2835..2851
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2835..2851
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2885..2901)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2886..2902)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3315..3334)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3316..3334)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3344..3361)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3344..3360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3353..3375)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3501..3956
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3588..3607)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3798..3819
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4087..4890)
                     /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(4891..5106)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5130..5149)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5249..5271
                     /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 gagctccacc
     2821 gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc gatatcaagc
     2881 ttatcgatac cgtcgacctc gagaacacaa cccacagcta ccaccatcaa caatatttat
     2941 atatataacg tacacataga aatcacacaa acagagtatt tattcttaac tacatgaact
     3001 accatcagac cgtctgggcc actatataat gtgccattca taaacgtgat cactttacgt
     3061 agcaggcaac cccaggtgaa aatttttcag cgagctgcca gattgtcagg tgaaaaactg
     3121 aaaaaaactt ctgggcgatg agcttgtggc gggaaattaa gtatataaag cagttagttt
     3181 cttctgcttc ttgtggttct gggtattctt gtaatatttg gaagaataga aaatcaagac
     3241 tactgccttt cttttcatat tatagaggaa gaaatacgca cgaacacgat atagaggtaa
     3301 aggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     3361 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3421 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3481 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3541 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3601 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3661 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3721 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3781 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3841 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3901 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     3961 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg gtaataactg
     4021 atataattaa attgaagctc taatttgtga gtttagtata catgcattta cttataatac
     4081 agttttttag ttttgctggc cgcatcttct caaatatgct tcccagcctg cttttctgta
     4141 acgttcaccc tctaccttag catcccttcc ctttgcaaat agtcctcttc caacaataat
     4201 aatgtcagat cctgtagaga ccacatcatc cacggttcta tactgttgac ccaatgcgtc
     4261 tcccttgtca tctaaaccca caccgggtgt cataatcaac caatcgtaac cttcatctct
     4321 tccacccatg tctctttgag caataaagcc gataacaaaa tctttgtcgc tcttcgcaat
     4381 gtcaacagta cccttagtat attctccagt agatagggag cccttgcatg acaattctgc
     4441 taacatcaaa aggcctctag gttcctttgt tacttcttct gccgcctgct tcaaaccgct
     4501 aacaatacct gggcccacca caccgtgtgc attcgtaatg tctgcccatt ctgctattct
     4561 gtatacaccc gcagagtact gcaatttgac tgtattacca atgtcagcaa attttctgtc
     4621 ttcgaagagt aaaaaattgt acttggcgga taatgccttt agcggcttaa ctgtgccctc
     4681 catggaaaaa tcagtcaaga tatccacatg tgtttttagt aaacaaattt tgggacctaa
     4741 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
     4801 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
     4861 agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt
     4921 tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
     4981 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
     5041 cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
     5101 attgaaaagg tggtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5161 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5221 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5281 tcatcaccga aacgcgcga
//
