LOCUS       sequence-106679-        4509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Initial replicative vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4509)
  AUTHORS   Yanez-Carrillo P, Orta-Zavalza E, Gutierrez-Escobedo G, 
            Patron-Soberano A, De Las Penas A, Castano I
  TITLE     Expression vectors for C-terminal fusions with fluorescent proteins 
            and epitope tags in Candida glabrata.
  JOURNAL   Fungal Genet Biol. 2015 May 15. pii: S1087-1845(15)00087-0. doi: 
            10.1016/j.fgb.2015.04.020.
  PUBMED    25986172
REFERENCE   2  (bases 1 to 4509)
  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"; journalName: "Fungal 
            Genet Biol. 2015 May 15. pii: S1087-1845(15)00087-0. doi: 
            10.1016/j.fgb.2015.04.020."
COMMENT     Sequence Label: pMJ22
FEATURES             Location/Qualifiers
     source          1..4509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        341..747
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             761..1333
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARELGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1340..1587
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1356..1374)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     misc_feature    1592..1699
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1608..1624
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1609..1625
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1659..1675)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1659..1675)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1712..1732)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1712..1730)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1751..1767)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1751..1767)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1764..1786)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1775..1791
                     /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(1799..1805,1806..1823,1824..1829))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1844..1865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1982..1999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2153..2741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2233..2252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2912..3703,3704..3772))
                     /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     3535..3554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3773..3844)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4482..4500
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcacctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca taccataaag cacagctcca tagcttcaaa
      361 atgtttctac tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca
      421 cttcaaaaca cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt
      481 taattacccg tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc
      541 gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt cttgaaaatt ttttttttga
      601 tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc
      661 tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag
      721 aaagaaagca tagcaatcta atctaagttt tggatctaaa atgggcacca ctcttgacga
      781 cacggcttac cgataccgca ccagtgtccc gggggacgcc gaggccatcg aggcactgga
      841 tgggtccttc accaccgaca ccgtcttccg cgtcaccgcc accggggacg gcttcaccct
      901 gcgggaggtg ccggtggacc cgcccctgac caaggtgttc cccgacgacg aatcggacga
      961 cgaatcggac gacggggagg acggcgaccc tgactcccgg acgttcgtcg cgtacgggga
     1021 cgacggcgac ctggcgggct tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac
     1081 cgtcgaggac atcgaggtcg ccccggagca ccgggggcac ggggtcgggc gcgcgttgat
     1141 ggggctcgcg acggagttcg cccgcgagct gggcgccggg cacctctggc tggaggtcac
     1201 caacgtcaac gcaccggcga tccacgcgta ccggcggatg gggttcaccc tctgcggcct
     1261 ggacaccgcc ctgtacgacg gcaccgcctc ggacggcgag caggcgctct acatgagcat
     1321 gccctgcccc taagtcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1381 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1441 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1501 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1561 ttgggacgct cgaaggcttt aatttgcggc cggtaccggg ccccccctcg aggtcgacgg
     1621 tatcgataag cttgatatcg aattcctgca gcccggggga tccactagtt ctagagcggc
     1681 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     1741 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1801 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1861 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1921 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1981 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2041 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2101 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2161 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2221 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2281 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2341 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2401 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2461 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2521 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2581 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2641 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2701 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2761 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2821 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2881 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2941 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3001 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3061 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3181 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3841 aatgtgggga tctctaatga tgcaattttt gttttgtaca ctaagattct atttcggaac
     3901 tcttacccat gtcattattt actttatttt cattattgta attttttaag cttttattat
     3961 gaattattta aatacaattc tttttgatca tgtgacagac atcactgaaa gatgtgctct
     4021 gcgttattag tcagattgaa atatacattt tcatgtagat atgatgttct tactttaaat
     4081 gttgttccat ttgctagctt aaatagacga tattataaat ttaagtattc actaaaaatc
     4141 tgacccatta atgttagaat tagtattatt ttttgtgata ttacattttt atattttgga
     4201 tttcaaccaa tgctctgcga tatgattcaa cagaataata tgtacaagct actttaattc
     4261 atatatacta atagattcac agatccagta cgactgcaag aatttacgtt ttgtttgaaa
     4321 tggtagtagc tacactaaat taacaatgat cgattaatcc acatccacag tgtgtaatca
     4381 tcctaatgaa aacaagaata actacaaaaa tgaatataaa ctttttgcat gttagtctaa
     4441 cctgctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4501 cctttcgtc
//
