LOCUS       sequence-242245-        7577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for tagging proteins in Cryptococcus neoformans; N-terminal 
            fragment of Venus protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7577)
  AUTHORS   So YS, Yang DH, Jung KW, Huh WK, Bahn YS
  TITLE     Molecular Characterization of Adenylyl Cyclase Complex Proteins 
            Using Versatile Protein-Tagging Plasmid Systems in Cryptococcus 
            neoformans.
  JOURNAL   J Microbiol Biotechnol. 2017 Feb 28;27(2):357-364. doi: 
            10.4014/jmb.1609.09036.
  PUBMED    27780958
REFERENCE   2  (bases 1 to 7577)
  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: "10.4014/jmb.1609"; 
            journalName: "J Microbiol Biotechnol"; date: "2017-02-28- 28"; 
            volume: "27"; issue: "2"; pages: "357-364"
COMMENT     Sequence Label: YSBE420:pNEO_CAC1_Vn_HOG1 terminator
FEATURES             Location/Qualifiers
     source          1..7577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1173..1175,1176..1688)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(1215..1236)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1476..1495)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(2313..3113)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MSMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4010..4029)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4011..4029)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4036..4053)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4036..4052)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4045..4067)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4193..4621
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4280..4299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4490..4511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             4965..5759
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(5777..5845,5846..6637)
                     /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(5995..6014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6808..7396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7297..7316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7550..7567
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatcccgt atgggtgttc attggggtgc gccagagtgc gaaaaagatc
      301 caatcacgag gaggatggac tactacggtc ccatggtgaa tcgtgctgcg cgtatcaatg
      361 caagtgcaga tggaggccag ttgatggcga gccaagacgt cctgaatgag attgcgccgc
      421 tgatggagta tctgaactca agtgatgaac aagtgttgaa tgatctgcag ggcgatttga
      481 agagggaagt tatggagttg agaagaattg gtctagaggt tagagacatg ggcgacagga
      541 agctcaaagg acttgaaggt gagcctggtc ttgttctgac tccatcaatg atatgctgac
      601 aatgcaacat agtgcccgaa aggcttcatt tgctttatcc caagactcta gccggccgac
      661 tggagatttc caacgagatc cgagccgaag tcgaagtcaa cgatgctcgc aaatctgctg
      721 aacgtcagcg cagtgttgat attgaccaag tctatcaact ttccgatatc gccttgcgat
      781 tagaagctgt ctgctgctac aaccctactc cctcatcacc tggtgacact cccacggcag
      841 gtgtcatgag attgcatcct ccagcaagtt atctggggcc ttctataaga gaggatatga
      901 acgatgaaga gctgtggacg atcattgaaa gcttagtggg gaggattgaa aacgtgatgt
      961 caactttggt gagtcgcggt catctaattt gttcataaat gcatgctgat cagccgttga
     1021 agtatctcaa aaactttgga gaatttagtg ctgtattggc cgctttggag tcggccacga
     1081 aaattgacca aaaactcatc gtgcatgcgt tggcgttaat gaacgaggcg atggggaaag
     1141 atgaagagaa cgcgataggt tctggtggta ctgtgagcaa gggcgaggag ctgttcaccg
     1201 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1261 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
     1321 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcctgcagt
     1381 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1441 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1501 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1561 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1621 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     1681 acatcgagta gggccgcgta aaagaatcgt tcttctttct ttctttcttt ttatttataa
     1741 cgaaagaaga gatgttggcg tatacttgaa cttatgaatg atgtgttgta ttcaggagcc
     1801 tgaaagtaat gtggtgtgtg gtgggcggtg ggacgaggaa gagtcgggat aagttgggat
     1861 gatagatggt ggagcaagat tgcataagca aaagttgaga agcgcaaagt tgagaagaag
     1921 tgtggacggc tcgtgttgcg ggttcaggga gcagaaggga gggaatgtgg agattgttcg
     1981 gagagaggga tagattggta cggatccact agtaacggcc gccagtgtgc tggaattcgc
     2041 ccttggttta tctgtattaa cacggaagag atgtagaaac tagcttcctg gtttcagaga
     2101 caaggagcca tgaagatcct gaggattcaa cggcgttgct gagggtggga aggatggtat
     2161 tgttgtctgt agatagatta atgggcatat aatgtcaaaa aagatgattg acacgatgtc
     2221 tgtctcgaat tcgcagtgtg ctaaaattgt atcacttctc taatatgtat atatacaccc
     2281 tctaaggaaa attaacccct taccgccttc actcagaaga actcgtcaag aaggcgatag
     2341 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     2401 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     2461 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     2521 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     2581 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     2641 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     2701 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     2761 atgatggata ctttctcggc aggagcaagg tgggatgaca ggagatcctg ccccggcact
     2821 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     2881 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     2941 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     3001 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     3061 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcataga catgttgggc
     3121 gagtttacta atggaaaaag acaattagca aagaggctgc atattaggag acgcggtgaa
     3181 gattggggag gacaataggg tccgtgcacc aaaaagtcct gacggcagct ctgttcaacg
     3241 cacatccatc tcttccagct atgcgtgtgg caccagcacg gatggattgt tgcaccaaaa
     3301 ggacaaggcg agatggatgc aggtcgaggc gatgcggtgc tatcggccct cgttgtcgaa
     3361 cgccacgtcg tcctctcggc gaccgtcaat cctggaagga aatgaccaag cgtgcaccca
     3421 gcatccagca tccaacatcc agcacccagc acctagcacc catgatccaa tggccggcat
     3481 gcagatcagc gtggggagag caccatctag cagcccctgg caggcccagc attgtgctcg
     3541 gatgtgcgat aggcggtggg tgcgataagg tggattagtg gaggatggag gactgagtgg
     3601 cgggcaaggg aagggggcgg gaagatgact caccttgggc agtgggtggg ggatgcggta
     3661 taggggatgt atgtggatgc tggcggagga tagaagctgt aagctgagct ggtgccaacg
     3721 gtgacgctgt gagagtggtc tcggcgaccc tgacggcgac tcggcgttac cccgtgtggc
     3781 acaccggacg cgcgtcgagt gacgttgctg ctattgttgt taccatcatc ctctcctccg
     3841 acaaccatac actcagtgca caccatgcca ctcgaatcct gcatgcttat gtgagtcctc
     3901 cccttcccgt gcgccgctct ccagctcaca tcctcgcagc aagggcgaat tctgcagata
     3961 tccatcacac tggcggccgc tcgagcatgc atctagaggg cccaattcgc cctatagtga
     4021 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     4081 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     4141 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     4201 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4261 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4321 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4381 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4441 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4501 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4561 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4621 tttaacaaaa ttcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     4681 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     4741 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     4801 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     4861 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     4921 ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4981 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5041 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5101 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5161 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5221 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatccc
     5281 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5341 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5401 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5461 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5521 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5581 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5641 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5701 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa
     5761 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5821 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5881 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5941 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6001 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6061 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     6121 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     6181 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6241 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6301 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     6361 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     6421 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6481 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6541 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6601 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     6661 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6721 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7441 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7501 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     7561 gagcgaggaa gcggaag
//
