LOCUS       sequence-242240-        5563 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 5563)
  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 5563)
  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: YSBE416: pTOP_CAC1_Vn_HOG1 terminator
FEATURES             Location/Qualifiers
     source          1..5563
                     /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             complement(join(614..1126,1127..1129))
                     /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     807..826
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1066..1087
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2146..2163)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2146..2162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2155..2177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2300..2602
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2481..2500
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2951..3745
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2951..3745
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3005..3024)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3615..3634
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3763..3831,3832..4623)
                     /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(3981..4000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4794..5382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5283..5302
                     /label=pBR322ori-F
                     /note="pBR322 origin, 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 cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttggatcc
      301 gtaccaatct atccctctct ccgaacaatc tccacattcc ctcccttctg ctccctgaac
      361 ccgcaacacg agccgtccac acttcttctc aactttgcgc ttctcaactt ttgcttatgc
      421 aatcttgctc caccatctat catcccaact tatcccgact cttcctcgtc ccaccgccca
      481 ccacacacca cattactttc aggctcctga atacaacaca tcattcataa gttcaagtat
      541 acgccaacat ctcttctttc gttataaata aaaagaaaga aagaaagaag aacgattctt
      601 ttacgcggcc ctactcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct
      661 gcttgtcggc ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
      721 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
      781 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
      841 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
      901 ggtcggggta gcgggcgaag cactgcaggc cgtagcccag ggtggtcacg agggtgggcc
      961 agggcacggg cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg
     1021 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1081 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcaca gtaccaccag
     1141 aacctatcgc gttctcttca tctttcccca tcgcctcgtt cattaacgcc aacgcatgca
     1201 cgatgagttt ttggtcaatt ttcgtggccg actccaaagc ggccaataca gcactaaatt
     1261 ctccaaagtt tttgagatac ttcaacggct gatcagcatg catttatgaa caaattagat
     1321 gaccgcgact caccaaagtt gacatcacgt tttcaatcct ccccactaag ctttcaatga
     1381 tcgtccacag ctcttcatcg ttcatatcct ctcttataga aggccccaga taacttgctg
     1441 gaggatgcaa tctcatgaca cctgccgtgg gagtgtcacc aggtgatgag ggagtagggt
     1501 tgtagcagca gacagcttct aatcgcaagg cgatatcgga aagttgatag acttggtcaa
     1561 tatcaacact gcgctgacgt tcagcagatt tgcgagcatc gttgacttcg acttcggctc
     1621 ggatctcgtt ggaaatctcc agtcggccgg ctagagtctt gggataaagc aaatgaagcc
     1681 tttcgggcac tatgttgcat tgtcagcata tcattgatgg agtcagaaca agaccaggct
     1741 caccttcaag tcctttgagc ttcctgtcgc ccatgtctct aacctctaga ccaattcttc
     1801 tcaactccat aacttccctc ttcaaatcgc cctgcagatc attcaacact tgttcatcac
     1861 ttgagttcag atactccatc agcggcgcaa tctcattcag gacgtcttgg ctcgccatca
     1921 actggcctcc atctgcactt gcattgatac gcgcagcacg attcaccatg ggaccgtagt
     1981 agtccatcct cctcgtgatt ggatcttttt cgcactctgg cgcaccccaa tgaacaccca
     2041 tacgggatcc aagggcgaat tctgcagata tccatcacac tggcggccgc tcgagcatgc
     2101 atctagaggg cccaattcgc cctatagtga atcgtattac aattcactgg ccgtcgtttt
     2161 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2221 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2281 gcgcagccta tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     2341 tctgtttgtg gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc
     2401 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     2461 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     2521 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     2581 cctgatgttc tggggaatat aaatgtcagg catgagatta tcaaaaagga tcttcaccta
     2641 gatccttttc acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag
     2701 ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag
     2761 tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt
     2821 gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt
     2881 cttgccgcca aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg
     2941 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3001 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3061 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3121 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3181 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3241 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3301 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3361 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3421 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3481 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3541 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3601 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3661 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3721 tcgccttctt gacgagttct tctgaattga aaaaggaaga gtatgagtat tcaacatttc
     3781 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3841 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3901 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3961 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4021 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4081 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4141 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4201 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4261 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4321 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4381 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4441 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4501 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagccaggca
     4561 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4621 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4681 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4741 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4861 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4921 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4981 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5041 ggcgataagt cgtgtcttac cgggttggat tcaagacgat agttaccgga taaggcgcag
     5101 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5161 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5221 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5281 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5341 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5401 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5461 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5521 cgaacgaccg agcgcagcga atcagtgagc gaggaagcgg aag
//
